Assay for measuring acetylation or deacetylation activity of an enzyme

ABSTRACT

This invention is directed to a continuous method for measuring the activity of an enzyme that catalyzes the addition of an acetyl group to a residue capable of being acetylated or an enzyme that catalyzes the removal of an acetyl group from an acetylated residue. In particular the present invention is directed to a continuous method for measuring the activity of histone acetyltranferases and histone deacetylase enzymes.

CROSS-REFERENCE

[0001] This application claims priority under 35 USC 119(e) to U.S.Provisional Application Serial No. 60/355,700, filed on Feb. 7, 2002,the disclosure of which is incorporated herein by reference in itsentirety.

FIELD OF THE INVENTION

[0002] This invention is directed to a continuous method for measuringthe activity of an enzyme that catalyzes the addition of an acetyl groupto a residue capable of being acetylated or an enzyme that catalyzes theremoval of an acetyl group from an acetylated residue. In particular thepresent invention is directed to a continuous method for measuring theactivity of histone acetyltranferases and histone deacetylase enzymes.

STATE OF THE ART

[0003] Acetylation and deacetylation of histone proteins, transcriptionfactors, and related proteins play a major role in the control ofcellular processes. In particular, the acetylation state of histonescontrols how tightly the histone proteins interact with DNA, andtherefore how accessible the DNA is to transcription factors. Enzymesthat add acetyl groups to histones or other proteins are called histoneacetyltransferases (HATs). Enzymes that remove the acetyl groups fallinto two families: the histone deacetylases (HDACs) and the Sir2 familyof deacetylases. Currently there are eleven known members of themammalian HDAC family (Gray and Ekström, Exper. Cell Res. 2001, 262,75-83; Zhou, et al. Proc. Natl. Acad. Sci. USA 2001, 98, 10572-10577;Kao et al. J. Biol. Chem. 2002, 277, 187-193; Gao et al. J. Biol. Chem.2002, 277, 25748-25755) and seven members of the Sir2 family (Gray andEkström, Exper. Cell Res. 2001, 262, 75-83).

[0004] Histone acetyltransferases catalyze the transfer of an acetylgroup from acetyl-CoA to the ε-amino group of a lysine residue on thetarget protein. Many HAT enzymes have been characterized from eukaryoticorganisms (Sterner and Berger, Microbiol. Mol. Biol. Rev. 2000, 64,435-459). HDAC enzymes utilize a zinc ion at the active site of theprotein to catalyze the removal of the acetyl group from acetyllysine inthe form of acetate. Members of the Sir2 family of enzymes use NAD as acofactor in the hydrolysis of acetyllysine.

[0005] The acetylation state of histone proteins plays a major role ingene expression and in cell-cycle control, and appears to play a role incertain forms of cancer. In particular, abnormal recruitment of histonedeacetylases by corepressor proteins has been shown to promote thedevelopment of promyelocytic leukemia. In tumor cell lines, severalstudies have shown that treatment with HDAC inhibitors can lead togrowth inhibition, growth arrest, terminal differentiation, and/orapoptosis. In vivo studies have demonstrated growth inhibition of tumorsand a reduction in tumor metastasis as a result of treatment with HDACinhibitors (Krämer et al. Trends Endocrinol. Metab. 2001, 12, 294-300).

[0006] Effective study of the enzymology and inhibition of HATs, HDACs,and Sir2 enzymes depends on the availability of robust assays capable ofbeing performed in a high-throughput manner. Several assay methodologieshave been developed for these enzymes, with varying degrees of utilityfor inhibitor screening.

[0007] Histone acetyltransferase assays are typicallyradioactivity-based. In these formats, acetyl-CoA radiolabeled on theacetyl group is reacted with a peptide corresponding to a histone aminoacid sequence. Transfer of radiolabeled acetate to the peptide isquantitated by binding of the peptide to affinity resin (Ait-Si-Ali etal. Nucleic Acids Res. 1998, 26, 3869-3870), phosphocellulose paper(Tanner et al. J. Biol. Chem. 1999, 274, 18157-18160), or scintillationmicroplates (Wynne Aherne et al. Methods 2002, 26, 245-53) andmeasurement of the associated radioactivity. In a non-radioactivecoupled assay format, the free CoA formed in the acetyltransferasereaction serves as a susbstrate for α-ketoglutarate dehydrogenase orpyruvate dehydrogenase. Formation of NADH serves as a measure of therate of acetyltransferase activity (Kim et al. Anal. Biochem. 2000, 280,308-314).

[0008] The most common deacetylase assay methodology involves labelinglysine groups in histone peptides with radiolabeled acetate. Thedeacetylase enzyme removes the acetyl group as acetate, which issubsequently isolated by extraction and quantitated on the basis of itsradioactivity (Inoue and Fujimoto, Biochim. Biophys. Acta 1970, 220,307-316). In a variant of this approach, a scintillation proximityassay, peptides derivatized with radiolabeled acetyl groups are attachedto a bead containing scintillant that emits light upon exposure toradiation. In this assay format, cleavage of the acetyl groups causes adecrease in the light emission from the scintillant (Nare, et al., Anal.Biochem. 1999, 267, 390-396). A non-radioactivity-based assay usespeptides containing an acetyllysine group and a fluorescent tag.Reactivity is measured by high-performance liquid chromatography, usingthe difference in retention time of the acetylated and non-acetylatedpeptides to isolate and quantitate the reaction products (Hoffmann etal. Nucleic Acids Res. 1999, 27, 2057-8; Hoffmann et al. Bioconjug Chem.2001,12, 51-5; Hoffmann et al. Arch Pharm (Weinheim) 2001, 334, 248-52).A commercial assay uses a two-step detection protocol. In the firststep, a peptide containing an acetyllysine is reacted with a deacetylasefor a given period of time. Following this, the reaction is quenched andthe exposed lysine is reacted with a developing agent that produces afluorophore, and the amount of deacetylated lysine is quantitated usingthe fluorescence of the product (Biomol, Plymouth Meeting, Pa., USA).More recently, a two-step, protease-coupled assay was reported, in whicha peptide was designed containing a fluorescence resonance energytransfer (FRET) donor-quencher pair and an acetyllysine. After thedeacetylase reaction has been allowed to run, the reaction is quenchedand the amount of of deacetylated peptide is quantitated by reaction ofthe deacetylated peptide with a protease enzyme that cleavesspecifically after lysine residues (Frey et al. Presented at 224thNational Meeting of the American Chemical Society, Boston, Mass., August2002; paper MEDI-121). To date, no continuous, non-radioactive histonedeacetylase assays have been reported.

[0009] Features of the above assay formats limit their utility. Assaysbased on radioactivity tend to be costly, and require special handlingprecautions. Also, they are often difficult to perform in ahigh-throughput manner. Assays that measure activity on the basis of thedisappearance of a signal with time rather that the appearance of signalusually yield poor signal/noise and signal/background ratios. Further,with the exception of the scintillation proximity assay described above,all deacetylase assays reported to date are endpoint assays, where theextent of the reaction is determined at a single time point only. Theendpoint format assumes a linear rate of product formation with time.However, this assumption is not valid if the enzyme follows burst-phaseor lag-phase kinetics, if the enzyme activity decreases over the courseof the reaction, if the substrate is significantly depleted over thecourse of the reaction, or if substrate or protein aggregation occursover the course of the reaction. A continous format, in which theprogress of the reaction is monitored as the reaction proceeds, is morerobust than an endpoint format.

[0010] Coupled assays are common in the practice of enzymology. Thetechnique has been reviewed in the literature (Rudolph, et al. MethodsEnz. 1979, 63, 22-42). In this type of assay, the formation of theproduct of an enzymatic reaction is not measured directly. Rather, theproduct reacts further with another enzyme or chemical to form a secondproduct that has is readily detectable using spectroscopic or otherdetection methodologies. Representative examples of this methodology area horseradish peroxidase-coupled assay for L-amino acid oxidase (Ueda etal. Toxicon 1988, 26, 695-706), a chymotrypsin-coupled assay for prolylisomerase (Fischer et al. Biomed. Biochim. Acta 1984, 43, 1101-11), andan assay for methionine aminopeptidase in which the cleavage of athioester bond in the enzyme substrate yields a free thiol that canreact with 5,5′-dithio-bis(2-nitrobenzoic acid) (DTNB) to yield adetectable chromophore (Zhou et al. Anal. Biochem. 2000, 280, 159-165).In a coupled enzyme system, if the second reaction is slower than thefirst, an endpoint format is necessary. The first reaction must bequenched, and the second reaction is used to quantify the amount ofproduct formed. In the case where the second reaction is more rapid,both reactions can be run at the same time. A steady-state can beachieved in which the rate of production of the first product will equalthe rate of its conversion into the second product. Under thiscondition, the signal produced by the formation of the second productwill be a measure of the formation of the first product, and thus ameasure of the activity of the first enzyme. This allows for acontinuous assay format, in which the enzymatic activity can bemonitored directly as a function of time.

SUMMARY OF THE INVENTION

[0011] In one aspect, this invention is directed to a continuous methodfor measuring the activity of an enzyme that catalyzes (a) the additionof an acetyl group to a residue capable of being acetylated or (b)removal of an acetyl group from an acetylated residue which methodcomprises incubating said enzyme with:

[0012] (i) a protease;

[0013] (ii) a polypeptide comprising:

[0014] (a) a recognition site for the protease;

[0015] (b) a residue, in which the acetylation state of the residuemodifies the rate of cleavage of the polypeptide by the protease; and

[0016] (c) at least one chemical moiety, attached to the polypeptide,that possesses an optical signal that changes upon cleavage of thepolypeptide; and

[0017] (iii) measuring the change in the optical signal.

[0018] In a second aspect, this invention is directed to a continuousmethod for measuring the activity of an enzyme that catalyzes theaddition of an acetyl group to a lysine residue or an enzyme thatcatalyzes the removal of acetyl group from N^(ε)-acetylated lysineresidue which method comprises incubating said enzyme with:

[0019] (i) a protease;

[0020] (ii) a polypeptide comprising:

[0021] (a) a recognition site for the protease;

[0022] (b) a lysine or acetyllysine residue, in which the acetylationstate of the residue modifies the rate of cleavage of the polypeptide bythe protease; and

[0023] (c) at least one chemical moiety, attached to the polypeptide,that possesses an optical signal that changes upon cleavage of thepolypeptide; and

[0024] (iii) measuring the change in the optical signal.

[0025] Preferably, the method measures the activity of an enzyme thatcatalyzes the removal of acetyl group from N^(ε)-acetylated lysine.Preferably, the enzyme is HDAC1 (SEQ. ID. NO: 1), HDAC2 (SEQ. ID. NO:2), HDAC3 (SEQ. ID. NO: 3), HDAC4 (SEQ. ID. NO: 4), HDAC5 (SEQ. ID. NO:5), HDAC6 (SEQ. ID. NO: 6), HDAC7 (SEQ. ID. NO: 7), HDAC8 (SEQ. ID. NO:8), HDAC9 (SEQ. ID. NO: 9), HDAC10 (SEQ. ID. NO: 10), or HDAC11 (SEQ.ID. NO: 11), any protein with 95% or greater sequence sirmilarity any ofthe said enzymes, or any fragment of any of the enzymes that retainscatalytic deacetylase activity. Preferably, the enzyme is SIRT1 (SEQ.ID. NO: 12), SIRT2 (SEQ. ID. NO: 13), SIRT3 (SEQ. ID. NO: 14), SIRT4(SEQ. ID. NO: 15), SIRT5 (SEQ. ID. NO: 16), SIRT6 (SEQ. ID. NO: 17), orSIRT7 (SEQ. ID. NO: 18), any protein with 95% or greater sequencesimilarity to any of the said enzymes, or any fragment of any of theenzymes that retains catalytic deacetylase activity. Most preferably,the enzyme is HDAC1.

[0026] Preferably, the protease is a member of the trypsin family ofproteases. More preferably, the protease is trypsin or thrombin.

[0027] Preferably, the optical signal arises from a fluorescent moiety,or optical absorption, or from a fluorescence resonance energy transfer.

[0028] Preferably, polypeptide is less than or equal to 8 or 20 aminoacids in length.

[0029] More preferably, the polypeptide isacetyl-Gly-Ala-(N^(ε)-acetyllysine)-AMC or(2-aminobenzoyl)-Gly-Ala-(N^(ε)-acetyllysine)-Ala-Ala-(3-(2,4-dinitrophenyl)-2,3-diaminopropionamide).Even more preferably acetyl-Gly-Ala-(N^(ε)-acetyllysine)-AMC.

[0030] In a third aspect, this invention is directed to a continuousmethod for measuring the inhibitory properties of a test compoundtowards the activity of an enzyme that catalyzes (a) the addition of anacetyl group to a residue capable of being acetylated or (b) removal ofan acetyl group from an acetylated residue which method comprisesincubating said enzyme with:

[0031] (i) a protease;

[0032] (ii) a polypeptide comprising:

[0033] (a) a recognition site for the protease;

[0034] (b) a residue, in which the acetylation state of the residuemodifies the rate of cleavage of the polypeptide by the protease; and

[0035] (c) at least one chemical moiety, attached to the polypeptide,that possesses an optical signal that changes upon cleavage of thepolypeptide;

[0036] in the presence of the test compound; and

[0037] (iii) measuring the rate of increase in the optical signalwherein the difference between the rate of increase of the opticalsignal in the presence of the test compound and the rate of increase ofthe optical signal in the absence of the test compound is indicative ofinhibitory properties of the test compound.

[0038] Preferably, this invention is directed to a continuous method formeasuring the inhibitory properties of a test compound towards an enzymethat catalyzes the addition or removal of acetyl group fromN^(ε)-acetylated lysine residue which method comprises incubating saidenzyme with:

[0039] (i) a protease;

[0040] (ii) a polypeptide comprising:

[0041] (a) a recognition site for the protease;

[0042] (b) a lysine or acetyllysine residue, in which the acetylationstate of the lysine residue modifies the rate of cleavage of thepolypeptide by the protease; and

[0043] (c) at least one chemical moiety, attached to the polypeptide,that possesses an optical signal that changes upon cleavage of thepolypeptide; in the presence of the test compound; and

[0044] (iii) measuring the rate of increase in the optical signalwherein the difference between the rate of increase of the opticalsignal in the presence of the test compound and the rate of increase ofthe optical signal in the absence of the test compound is indicative ofinhibitory properties of the test compound.

[0045] More preferably, the invention is directed to a continuous methodfor measuring the inhibitory properties of a test compound towards ahistone deacetylase enzyme which method comprises incubating the histonedeacetylase enzyme with:

[0046] (i) trypsin;

[0047] (ii) acetyl-Gly-Ala-(N^(ε)-acetyllysine)-AMC; in the presence andabsence of the test compound; and

[0048] (iii) measuring the increase in fluorescence at 460 nm over time,using an excitation wavelength of 355 nm wherein the difference in therate of increase of the fluorescence in the presence and absence of thetest compound is indicative of inhibitory properties of the testcompound.

[0049] Even more preferably, the histone deacetylase enzyme is HDAC1.

[0050] Particularly preferably, HDAC1 is incubated with the testcompound for at least 5 minutes prior to addition of trypsin andacetyl-Gly-Ala-(N^(ε)-acetyllysine)-AMC.

Definitions

[0051] Unless otherwise stated, the following terms used in thespecification and claims are defined for the purposes of thisApplication and have the following meanings. Other terms used in thespecification and claims have meanings recognized in the art.

[0052] The term “a” as used herein means at least one.

[0053] A “polypeptide” or “peptide” as used herein is a sequence ofamino acids joined through amide bonds. The amino acids may be naturallyoccurring or non-natural. It is known in the art that side chains ofseveral naturally occurring amino acids may be modified by the additionof chemical functionalities comprising methyl, acetyl, or phosphategroups.

[0054] A “moiety” as used herein is a molecule or portion of a moleculethat possesses an optical signal or imparts an optical property to themolecule to which it is bound. For example, the amide form of7-amino-4-methylcoumarin (AMC) when bound to a polypeptide has a weakfluorescence with an emission maximum at 395 nm. When the amide bond iscleaved, the free 7-amino-4-methylcoumarin has a very high fluorescenceintensity with an emission maximum at 460 nm. Similarly, thep-nitroaniline moiety in its amide form is colorless, but when cleavedfrom the polypeptide it acquires an intense yellow color.

[0055] A “continuous” assay or method as used herein is one in which theprocess can be monitored on a constant basis without changing theprocess as a result of the measurement. This includes any technique inwhich the sample is monitored multiple times over the course of thereaction, but the preferred definition is one in which formation of adetectable product is directly related to the activity of the enzyme andthe product can be quantitated in situ without any additional liquidhandling or chemical reaction steps. For example, in the HDAC assayinvolving acetyl-Gly-Ala-(N^(ε)-acetyllysine)-AMC, the rate ofproduction of free AMC is a measure of the rate of the deacetylasereaction. The production of free AMC can be monitored constantly or atarbitrarily small time intervals on the basis of its fluorescence, andthus the assay method is continuous. The reaction does not have to bestopped to detect a signal, and there are no extraction or purificationsteps necessary to isolate and quantitate the products.

[0056] A “proteolytic enzyme” or “protease” as used herein is an enzymethat catalyzes the cleavage of amide bonds within a polypeptide. Trypsinand thrombin are examples of proteases.

[0057] An “optical signal” as used herein is any response toillumination of a moiety that can be used to detect or quantitate thegiven moiety. Absorption and fluorescence are examples of opticalsignals.

[0058] A “residue” as used herein in the context of a polypeptide is anamino acid side chain that occurs within the polypeptide chain. Theresidue can be naturally or non-naturally occurring. For example,hydroxymethyl, thiomethyl, are naturally occurring residues.

[0059] A “recognition site” as used herein is a sequence of amino acidswithin a polypeptide that allows a protease enzyme to bind to and cleavethe said polypeptide.

[0060] Unless otherwise stated, “acetyllysine” as used herein refers tolysine acetylated at the ε-amino nitrogen.

[0061] “Inhibition” as used herein is a decrease in the rate of anenzyme-catalyzed reaction as a result of a compound binding to theenzyme and disrupting the interaction of the enzyme with its substrate.For the assays described herein, inhibition of a deacetylase enzyme willresult in a decrease in the magnitude of the optical signal as comparedwith a reaction in the absence of an inhibitor.

EXAMPLES Example 1 Synthesis of acetyl-Gly-Ala-(N-acetyl-Lys)-AMC

[0062] tert-Boc (N-Acetyl-Lys)-AMC (445 mg, 1 mmol, purchased fromBachem) was dissolved in 4 M HCL in dioxane to provideH-(N-acetyl-Lys)-AMC as a white solid. To a solution ofH-(N-acetyl-Lys)-AMC in DMF (5 ml) was added Ac-Gly-Ala-OH (188 mg, 1mmol) using PyBOP (520 mg, 1 mmol), HOBt (135 mg, 1 mmol), and NMM(0.296 ml, 2 mmol). The reaction mixture was stirred for 1 h andmonitored by MS/LC for the presence of H-(N-acetyl-Lys)-AMC. Additionalamounts of PyBOP (260 mg, 0.5 mmol), HOBt (70 mg, 0.5 mmol), and NMM(0.146 ml, 1 mmol) was added and the stirring was continued foradditional 4 h after which the product was isolated in quantative yield.

Example 2

[0063] Measurement of Histone Deacetylase Activity Using a FRETSubstrate

[0064] HDAC8 was cloned, isolated, and purified as described in theliterature (Buggy, et al. Biochem. J. 2000, 350, 199-205). The peptide2-aminobenzoyl-Gly-Ala-(N^(ε)-acetyllysine)-Ala-Ala-(3-dinitrophenyl-(L)-2,3-diaminopropionamide)(peptide 2) was purchased from California Peptide Research, Inc. Themeasurement was performed in a reaction volume of 100 μL using a 96-wellassay plate. HDAC8 (approx. 400 nM final concentration) in 50 mM HEPES,100 mM KCl, 0.001% Tween-20, 0.01% bovine serum albumin, 5% DMSO, pH7.4, was mixed with bovine trypsin (Sigma, 50 nM final concentration)and peptide 2 (20 μM final concentration). The reaction was monitoredfor 1 hour in a fluorescence plate reader, using an excitationwavelength of 320 nm and a detection wavelength of 405 nm. An increaseof fluorescence with time was used as the measure of reaction rate.

Example 3 Determination of the Inhibitory Properties of ChemicalCompounds

[0065] Measurements were performed in a reaction volume of 100 μL using96-well assay plates. HDAC-1 (200 pM final concentration) in reactionbuffer (50 mM HEPES, 100 mM KCl, 0.001% Tween-20, 5% DMSO, pH 7.4) wasmixed with inhibitor at various concentrations and allowed to incubatefor 30 minutes, after which trypsin andacetyl-Gly-Ala-(N-acetyl-Lys)-AMC were added to final concentrations of50 nM and 25 μM, respectively, to initiate the reaction. Negativecontrol reactions were performed in the absence of inhibitor inreplicates of eight.

[0066] The reactions were monitored in a fluorescence plate reader.After a 30 minute lag time, the fluorescence was measured over a 30minute time frame using an excitation wavelength of 355 nm and adetection wavelength of 460 nm. The increase in fluorescence with timewas used as the measure of the reaction rate. Inhibition constants wereobtained using the program BatchKi (Kuzmic et al. Anal. Biochem. 2000,286, 45-50).

Sequence Identification

[0067] Sequence IDs utiltize the accession codes of the NCBI ReferenceSequence project at the National Institutes of Health. Supportingliterature references are included.

[0068] SEQ ID. NO: 1 (Taunton, et al. Science 1996, 272, 408-411; RefSeqNP_(—)004955)

[0069] SEQ ID. NO: 2 (Yang, et al. Proc. Natl. Acad. Sci. U.S.A. 1996,93, 12845-12850; RefSeq NP_(—)001518)

[0070] SEQ ID. NO: 3 (Yang et al. J. Biol. Chem. 1997, 272, 28001-28007;RefSeq NP_(—)003874)

[0071] SEQ ID. NO: 4 (Grozinger, et al. Proc. Natl. Acad. Sci. U.S.A.1999, 96, 4868-4873; RefSeq NP_(—)006028)

[0072] SEQ ID. NO: 5 (Grozinger, et al. Proc. Natl. Acad. Sci. U.S.A.1999, 96, 4868-4873; RefSeq NP_(—)631944)

[0073] SEQ ID NO: 6 (Grozinger, et al. Proc. Natl. Acad. Sci. U.S.A.1999, 96, 4868-4873; RefSeq NP_(—)006035)

[0074] SEQ ID. NO: 7 (Kao, et al. Genes Dev. 2000, 14, 55-66; RefSeqNP_(—)056216)

[0075] SEQ ID. NO: 8 (Buggy, et al. Biochem. J. 2000, 350, 199-205;RefSeq NP_(—)060956)

[0076] SEQ ID. NO: 9 (Zhou, et al. Proc. Natl. Acad. Sci. U.S.A. 2001,98, 10572-10577; RefSeq NP_(—)478056)

[0077] SEQ ID. NO: 10 (Kao, et al. J. Biol. Chem. 2002, 277, 187-193;RefSeq NP_(—)114408)

[0078] SEQ ID. NO: 11 (Gao, et al. J. Biol. Chem. 2002, 277,25748-25755; RefSeq NP_(—)079103)

[0079] SEQ ID. NO: 12 (Frye, Biochem. Biophys. Res. Commun. 1999, 260,273-279; RefSeq NP_(—)036370)

[0080] SEQ ID. NO: 13 (Frye, Biochem. Biophys. Res. Commun. 1999, 260,273-279; RefSeq NP_(—)036369)

[0081] SEQ ID. NO: 14 (Frye, Biochem. Biophys. Res. Commun. 1999, 260,273-279; RefSeq NP_(—)036371)

[0082] SEQ ID. NO: 15 (Frye, Biochem. Biophys. Res. Commun. 1999, 260,273-279; RefSeq NP_(—)036372)

[0083] SEQ ID. NO: 16 (Frye, Biochem. Biophys. Res. Commun. 1999, 260,273-279; RefSeq NP_(—)036373)

[0084] SEQ ID. NO: 17 (Frye, Biochem. Biophys. Res. Commun. 2000, 273,793-798; RefSeq NP_(—)057623)

[0085] SEQ ID NO: 18 (Frye, Biochem. Biophys. Res. Commun. 2000, 273,793-798; RefSeq NP_(—)057622)

[0086] The foregoing invention has been described in some detail by wayof illustration and example, for purposes of clarity and understanding.It will be obvious to one of skill in the art that changes andmodifications may be practiced within the scope of the appended claims.Therefore, it is to be understood that the above description is intendedto be illustrative and not restrictive. The scope of the inventionshould, therefore, be determined not with reference to the abovedescription, but should instead be determined with reference to thefollowing appended claims, along with the full scope of equivalents towhich such claims are entitled. All patents, patent applications andpublications cited in this application are hereby incorporated byreference in their entirety for all purposes to the same extent as ifeach individual patent, patent application or publication were soindividually denoted.

1 18 1 482 PRT Homo sapiens 1 Met Ala Gln Thr Gln Gly Thr Arg Arg LysVal Cys Tyr Tyr Tyr Asp 1 5 10 15 Gly Asp Val Gly Asn Tyr Tyr Tyr GlyGln Gly His Pro Met Lys Pro 20 25 30 His Arg Ile Arg Met Thr His Asn LeuLeu Leu Asn Tyr Gly Leu Tyr 35 40 45 Arg Lys Met Glu Ile Tyr Arg Pro HisLys Ala Asn Ala Glu Glu Met 50 55 60 Thr Lys Tyr His Ser Asp Asp Tyr IleLys Phe Leu Arg Ser Ile Arg 65 70 75 80 Pro Asp Asn Met Ser Glu Tyr SerLys Gln Met Gln Arg Phe Asn Val 85 90 95 Gly Glu Asp Cys Pro Val Phe AspGly Leu Phe Glu Phe Cys Gln Leu 100 105 110 Ser Thr Gly Gly Ser Val AlaSer Ala Val Lys Leu Asn Lys Gln Gln 115 120 125 Thr Asp Ile Ala Val AsnTrp Ala Gly Gly Leu His His Ala Lys Lys 130 135 140 Ser Glu Ala Ser GlyPhe Cys Tyr Val Asn Asp Ile Val Leu Ala Ile 145 150 155 160 Leu Glu LeuLeu Lys Tyr His Gln Arg Val Leu Tyr Ile Asp Ile Asp 165 170 175 Ile HisHis Gly Asp Gly Val Glu Glu Ala Phe Tyr Thr Thr Asp Arg 180 185 190 ValMet Thr Val Ser Phe His Lys Tyr Gly Glu Tyr Phe Pro Gly Thr 195 200 205Gly Asp Leu Arg Asp Ile Gly Ala Gly Lys Gly Lys Tyr Tyr Ala Val 210 215220 Asn Tyr Pro Leu Arg Asp Gly Ile Asp Asp Glu Ser Tyr Glu Ala Ile 225230 235 240 Phe Lys Pro Val Met Ser Lys Val Met Glu Met Phe Gln Pro SerAla 245 250 255 Val Val Leu Gln Cys Gly Ser Asp Ser Leu Ser Gly Asp ArgLeu Gly 260 265 270 Cys Phe Asn Leu Thr Ile Lys Gly His Ala Lys Cys ValGlu Phe Val 275 280 285 Lys Ser Phe Asn Leu Pro Met Leu Met Leu Gly GlyGly Gly Tyr Thr 290 295 300 Ile Arg Asn Val Ala Arg Cys Trp Thr Tyr GluThr Ala Val Ala Leu 305 310 315 320 Asp Thr Glu Ile Pro Asn Glu Leu ProTyr Asn Asp Tyr Phe Glu Tyr 325 330 335 Phe Gly Pro Asp Phe Lys Leu HisIle Ser Pro Ser Asn Met Thr Asn 340 345 350 Gln Asn Thr Asn Glu Tyr LeuGlu Lys Ile Lys Gln Arg Leu Phe Glu 355 360 365 Asn Leu Arg Met Leu ProHis Ala Pro Gly Val Gln Met Gln Ala Ile 370 375 380 Pro Glu Asp Ala IlePro Glu Glu Ser Gly Asp Glu Asp Glu Asp Asp 385 390 395 400 Pro Asp LysArg Ile Ser Ile Cys Ser Ser Asp Lys Arg Ile Ala Cys 405 410 415 Glu GluGlu Phe Ser Asp Ser Glu Glu Glu Gly Glu Gly Gly Arg Lys 420 425 430 AsnSer Ser Asn Phe Lys Lys Ala Lys Arg Val Lys Thr Glu Asp Glu 435 440 445Lys Glu Lys Asp Pro Glu Glu Lys Lys Glu Val Thr Glu Glu Glu Lys 450 455460 Thr Lys Glu Glu Lys Pro Glu Ala Lys Gly Val Lys Glu Glu Val Lys 465470 475 480 Leu Ala 2 488 PRT Homo sapiens 2 Met Ala Tyr Ser Gln Gly GlyGly Lys Lys Lys Val Cys Tyr Tyr Tyr 1 5 10 15 Asp Gly Asp Ile Gly AsnTyr Tyr Tyr Gly Gln Gly His Pro Met Lys 20 25 30 Pro His Arg Ile Arg MetThr His Asn Leu Leu Leu Asn Tyr Gly Leu 35 40 45 Tyr Arg Lys Met Glu IleTyr Arg Pro His Lys Ala Thr Ala Glu Glu 50 55 60 Met Thr Lys Tyr His SerAsp Glu Tyr Ile Lys Phe Leu Arg Ser Ile 65 70 75 80 Arg Pro Asp Asn MetSer Glu Tyr Ser Lys Gln Met His Ile Phe Asn 85 90 95 Val Gly Glu Asp CysPro Ala Phe Asp Gly Leu Phe Glu Phe Cys Gln 100 105 110 Leu Ser Thr GlyGly Ser Val Ala Gly Ala Val Lys Leu Asn Arg Gln 115 120 125 Gln Thr AspMet Ala Val Asn Trp Ala Gly Gly Leu His His Ala Lys 130 135 140 Lys TyrGlu Ala Ser Gly Phe Cys Tyr Val Asn Asp Ile Val Leu Ala 145 150 155 160Ile Leu Glu Leu Leu Lys Tyr His Gln Arg Val Leu Tyr Ile Asp Ile 165 170175 Asp Ile His His Gly Asp Gly Val Glu Glu Ala Phe Tyr Thr Thr Asp 180185 190 Arg Val Met Thr Val Ser Phe His Lys Tyr Gly Glu Tyr Phe Pro Gly195 200 205 Thr Gly Asp Leu Arg Asp Ile Gly Ala Gly Lys Gly Lys Tyr TyrAla 210 215 220 Val Asn Phe Pro Met Cys Asp Gly Ile Asp Asp Glu Ser TyrGly Gln 225 230 235 240 Ile Phe Lys Pro Ile Ile Ser Lys Val Met Glu MetTyr Gln Pro Ser 245 250 255 Ala Val Val Leu Gln Cys Gly Ala Asp Ser LeuSer Gly Asp Arg Leu 260 265 270 Gly Cys Phe Asn Leu Thr Val Lys Gly HisAla Lys Cys Val Glu Val 275 280 285 Val Lys Thr Phe Asn Leu Pro Leu LeuMet Leu Gly Gly Gly Gly Tyr 290 295 300 Thr Ile Arg Asn Val Ala Arg CysTrp Thr Tyr Glu Thr Ala Val Ala 305 310 315 320 Leu Asp Cys Glu Ile ProAsn Glu Leu Pro Tyr Asn Asp Tyr Phe Glu 325 330 335 Tyr Phe Gly Pro AspPhe Lys Leu His Ile Ser Pro Ser Asn Met Thr 340 345 350 Asn Gln Asn ThrPro Glu Tyr Met Glu Lys Ile Lys Gln Arg Leu Phe 355 360 365 Glu Asn LeuArg Met Leu Pro His Ala Pro Gly Val Gln Met Gln Ala 370 375 380 Ile ProGlu Asp Ala Val His Glu Asp Ser Gly Asp Glu Asp Gly Glu 385 390 395 400Asp Pro Asp Lys Arg Ile Ser Ile Arg Ala Ser Asp Lys Arg Ile Ala 405 410415 Cys Asp Glu Glu Phe Ser Asp Ser Glu Asp Glu Gly Glu Gly Gly Arg 420425 430 Arg Asn Val Ala Asp His Lys Lys Gly Ala Lys Lys Ala Arg Ile Glu435 440 445 Glu Asp Lys Lys Glu Thr Glu Asp Lys Lys Thr Asp Val Lys GluGlu 450 455 460 Asp Lys Ser Lys Asp Asn Ser Gly Glu Lys Thr Asp Thr LysGly Thr 465 470 475 480 Lys Ser Glu Gln Leu Ser Asn Pro 485 3 428 PRTHomo sapiens 3 Met Ala Lys Thr Val Ala Tyr Phe Tyr Asp Pro Asp Val GlyAsn Phe 1 5 10 15 His Tyr Gly Ala Gly His Pro Met Lys Pro His Arg LeuAla Leu Thr 20 25 30 His Ser Leu Val Leu His Tyr Gly Leu Tyr Lys Lys MetIle Val Phe 35 40 45 Lys Pro Tyr Gln Ala Ser Gln His Asp Met Cys Arg PheHis Ser Glu 50 55 60 Asp Tyr Ile Asp Phe Leu Gln Arg Val Ser Pro Thr AsnMet Gln Gly 65 70 75 80 Phe Thr Lys Ser Leu Asn Ala Phe Asn Val Gly AspAsp Cys Pro Val 85 90 95 Phe Pro Gly Leu Phe Glu Phe Cys Ser Arg Tyr ThrGly Ala Ser Leu 100 105 110 Gln Gly Ala Thr Gln Leu Asn Asn Lys Ile CysAsp Ile Ala Ile Asn 115 120 125 Trp Ala Gly Gly Leu His His Ala Lys LysPhe Glu Ala Ser Gly Phe 130 135 140 Cys Tyr Val Asn Asp Ile Val Ile GlyIle Leu Glu Leu Leu Lys Tyr 145 150 155 160 His Pro Arg Val Leu Tyr IleAsp Ile Asp Ile His His Gly Asp Gly 165 170 175 Val Gln Glu Ala Phe TyrLeu Thr Asp Arg Val Met Thr Val Ser Phe 180 185 190 His Lys Tyr Gly AsnTyr Phe Phe Pro Gly Thr Gly Asp Met Tyr Glu 195 200 205 Val Gly Ala GluSer Gly Arg Tyr Tyr Cys Leu Asn Val Pro Leu Arg 210 215 220 Asp Gly IleAsp Asp Gln Ser Tyr Lys His Leu Phe Gln Pro Val Ile 225 230 235 240 AsnGln Val Val Asp Phe Tyr Gln Pro Thr Cys Ile Val Leu Gln Cys 245 250 255Gly Ala Asp Ser Leu Gly Cys Asp Arg Leu Gly Cys Phe Asn Leu Ser 260 265270 Ile Arg Gly His Gly Glu Cys Val Glu Tyr Val Lys Ser Phe Asn Ile 275280 285 Pro Leu Leu Val Leu Gly Gly Gly Gly Tyr Thr Val Arg Asn Val Ala290 295 300 Arg Cys Trp Thr Tyr Glu Thr Ser Leu Leu Val Glu Glu Ala IleSer 305 310 315 320 Glu Glu Leu Pro Tyr Ser Glu Tyr Phe Glu Tyr Phe AlaPro Asp Phe 325 330 335 Thr Leu His Pro Asp Val Ser Thr Arg Ile Glu AsnGln Asn Ser Arg 340 345 350 Gln Tyr Leu Asp Gln Ile Arg Gln Thr Ile PheGlu Asn Leu Lys Met 355 360 365 Leu Asn His Ala Pro Ser Val Gln Ile HisAsp Val Pro Ala Asp Leu 370 375 380 Leu Thr Tyr Asp Arg Thr Asp Glu AlaAsp Ala Glu Glu Arg Gly Pro 385 390 395 400 Glu Glu Asn Tyr Ser Arg ProGlu Ala Pro Asn Glu Phe Tyr Asp Gly 405 410 415 Asp His Asp Asn Asp LysGlu Ser Asp Val Glu Ile 420 425 4 1084 PRT Homo sapiens 4 Met Ser SerGln Ser His Pro Asp Gly Leu Ser Gly Arg Asp Gln Pro 1 5 10 15 Val GluLeu Leu Asn Pro Ala Arg Val Asn His Met Pro Ser Thr Val 20 25 30 Asp ValAla Thr Ala Leu Pro Leu Gln Val Ala Pro Ser Ala Val Pro 35 40 45 Met AspLeu Arg Leu Asp His Gln Phe Ser Leu Pro Val Ala Glu Pro 50 55 60 Ala LeuArg Glu Gln Gln Leu Gln Gln Glu Leu Leu Ala Leu Lys Gln 65 70 75 80 LysGln Gln Ile Gln Arg Gln Ile Leu Ile Ala Glu Phe Gln Arg Gln 85 90 95 HisGlu Gln Leu Ser Arg Gln His Glu Ala Gln Leu His Glu His Ile 100 105 110Lys Gln Gln Gln Glu Met Leu Ala Met Lys His Gln Gln Glu Leu Leu 115 120125 Glu His Gln Arg Lys Leu Glu Arg His Arg Gln Glu Gln Glu Leu Glu 130135 140 Lys Gln His Arg Glu Gln Lys Leu Gln Gln Leu Lys Asn Lys Glu Lys145 150 155 160 Gly Lys Glu Ser Ala Val Ala Ser Thr Glu Val Lys Met LysLeu Gln 165 170 175 Glu Phe Val Leu Asn Lys Lys Lys Ala Leu Ala His ArgAsn Leu Asn 180 185 190 His Cys Ile Ser Ser Asp Pro Arg Tyr Trp Tyr GlyLys Thr Gln His 195 200 205 Ser Ser Leu Asp Gln Ser Ser Pro Pro Gln SerGly Val Ser Thr Ser 210 215 220 Tyr Asn His Pro Val Leu Gly Met Tyr AspAla Lys Asp Asp Phe Pro 225 230 235 240 Leu Arg Lys Thr Ala Ser Glu ProAsn Leu Lys Leu Arg Ser Arg Leu 245 250 255 Lys Gln Lys Val Ala Glu ArgArg Ser Ser Pro Leu Leu Arg Arg Lys 260 265 270 Asp Gly Pro Val Val ThrAla Leu Lys Lys Arg Pro Leu Asp Val Thr 275 280 285 Asp Ser Ala Cys SerSer Ala Pro Gly Ser Gly Pro Ser Ser Pro Asn 290 295 300 Asn Ser Ser GlySer Val Ser Ala Glu Asn Gly Ile Ala Pro Ala Val 305 310 315 320 Pro SerIle Pro Ala Glu Thr Ser Leu Ala His Arg Leu Val Ala Arg 325 330 335 GluGly Ser Ala Ala Pro Leu Pro Leu Tyr Thr Ser Pro Ser Leu Pro 340 345 350Asn Ile Thr Leu Gly Leu Pro Ala Thr Gly Pro Ser Ala Gly Thr Ala 355 360365 Gly Gln Gln Asp Thr Glu Arg Leu Thr Leu Pro Ala Leu Gln Gln Arg 370375 380 Leu Ser Leu Phe Pro Gly Thr His Leu Thr Pro Tyr Leu Ser Thr Ser385 390 395 400 Pro Leu Glu Arg Asp Gly Gly Ala Ala His Ser Pro Leu LeuGln His 405 410 415 Met Val Leu Leu Glu Gln Pro Pro Ala Gln Ala Pro LeuVal Thr Gly 420 425 430 Leu Gly Ala Leu Pro Leu His Ala Gln Ser Leu ValGly Ala Asp Arg 435 440 445 Val Ser Pro Ser Ile His Lys Leu Arg Gln HisArg Pro Leu Gly Arg 450 455 460 Thr Gln Ser Ala Pro Leu Pro Gln Asn AlaGln Ala Leu Gln His Leu 465 470 475 480 Val Ile Gln Gln Gln His Gln GlnPhe Leu Glu Lys His Lys Gln Gln 485 490 495 Phe Gln Gln Gln Gln Leu GlnMet Asn Lys Ile Ile Pro Lys Pro Ser 500 505 510 Glu Pro Ala Arg Gln ProGlu Ser His Pro Glu Glu Thr Glu Glu Glu 515 520 525 Leu Arg Glu His GlnAla Leu Leu Asp Glu Pro Tyr Leu Asp Arg Leu 530 535 540 Pro Gly Gln LysGlu Ala His Ala Gln Ala Gly Val Gln Val Lys Gln 545 550 555 560 Glu ProIle Glu Ser Asp Glu Glu Glu Ala Glu Pro Pro Arg Glu Val 565 570 575 GluPro Gly Gln Arg Gln Pro Ser Glu Gln Glu Leu Leu Phe Arg Gln 580 585 590Gln Ala Leu Leu Leu Glu Gln Gln Arg Ile His Gln Leu Arg Asn Tyr 595 600605 Gln Ala Ser Met Glu Ala Ala Gly Ile Pro Val Ser Phe Gly Gly His 610615 620 Arg Pro Leu Ser Arg Ala Gln Ser Ser Pro Ala Ser Ala Thr Phe Pro625 630 635 640 Val Ser Val Gln Glu Pro Pro Thr Lys Pro Arg Phe Thr ThrGly Leu 645 650 655 Val Tyr Asp Thr Leu Met Leu Lys His Gln Cys Thr CysGly Ser Ser 660 665 670 Ser Ser His Pro Glu His Ala Gly Arg Ile Gln SerIle Trp Ser Arg 675 680 685 Leu Gln Glu Thr Gly Leu Arg Gly Lys Cys GluCys Ile Arg Gly Arg 690 695 700 Lys Ala Thr Leu Glu Glu Leu Gln Thr ValHis Ser Glu Ala His Thr 705 710 715 720 Leu Leu Tyr Gly Thr Asn Pro LeuAsn Arg Gln Lys Leu Asp Ser Lys 725 730 735 Lys Leu Leu Gly Ser Leu AlaSer Val Phe Val Arg Leu Pro Cys Gly 740 745 750 Gly Val Gly Val Asp SerAsp Thr Ile Trp Asn Glu Val His Ser Ala 755 760 765 Gly Ala Ala Arg LeuAla Val Gly Cys Val Val Glu Leu Val Phe Lys 770 775 780 Val Ala Thr GlyGlu Leu Lys Asn Gly Phe Ala Val Val Arg Pro Pro 785 790 795 800 Gly HisHis Ala Glu Glu Ser Thr Pro Met Gly Phe Cys Tyr Phe Asn 805 810 815 SerVal Ala Val Ala Ala Lys Leu Leu Gln Gln Arg Leu Ser Val Ser 820 825 830Lys Ile Leu Ile Val Asp Trp Asp Val His His Gly Asn Gly Thr Gln 835 840845 Gln Ala Phe Tyr Ser Asp Pro Ser Val Leu Tyr Met Ser Leu His Arg 850855 860 Tyr Asp Asp Gly Asn Phe Phe Pro Gly Ser Gly Ala Pro Asp Glu Val865 870 875 880 Gly Thr Gly Pro Gly Val Gly Phe Asn Val Asn Met Ala PheThr Gly 885 890 895 Gly Leu Asp Pro Pro Met Gly Asp Ala Glu Tyr Leu AlaAla Phe Arg 900 905 910 Thr Val Val Met Pro Ile Ala Ser Glu Phe Ala ProAsp Val Val Leu 915 920 925 Val Ser Ser Gly Phe Asp Ala Val Glu Gly HisPro Thr Pro Leu Gly 930 935 940 Gly Tyr Asn Leu Ser Ala Arg Cys Phe GlyTyr Leu Thr Lys Gln Leu 945 950 955 960 Met Gly Leu Ala Gly Gly Arg IleVal Leu Ala Leu Glu Gly Gly His 965 970 975 Asp Leu Thr Ala Ile Cys AspAla Ser Glu Ala Cys Val Ser Ala Leu 980 985 990 Leu Gly Asn Glu Leu AspPro Leu Pro Glu Lys Val Leu Gln Gln Arg 995 1000 1005 Pro Asn Ala AsnAla Val Arg Ser Met Glu Lys Val Met Glu Ile His 1010 1015 1020 Ser LysTyr Trp Arg Cys Leu Gln Arg Thr Thr Ser Thr Ala Gly Arg 1025 1030 10351040 Ser Leu Ile Glu Ala Gln Thr Cys Glu Asn Glu Glu Ala Glu Thr Val1045 1050 1055 Thr Ala Met Ala Ser Leu Ser Val Gly Val Lys Pro Ala GluLys Arg 1060 1065 1070 Pro Asp Glu Glu Pro Met Glu Glu Glu Pro Pro Leu1075 1080 5 1037 PRT Homo sapiens 5 Met Asn Ser Pro Asn Glu Ser Asp GlyMet Ser Gly Arg Glu Pro Ser 1 5 10 15 Leu Glu Ile Leu Pro Arg Thr SerLeu His Ser Ile Pro Val Thr Val 20 25 30 Glu Val Lys Pro Val Leu Pro ArgAla Met Pro Ser Ser Met Gly Gly 35 40 45 Gly Gly Gly Gly Ser Pro Ser ProVal Glu Leu Arg Gly Ala Leu Val 50 55 60 Gly Ser Val Asp Pro Thr Leu ArgGlu Gln Gln Leu Gln Gln Glu Leu 65 70 75 80 Leu Ala Leu Lys Gln Gln GlnGln Leu Gln Lys Gln Leu Leu Phe Ala 85 90 95 Glu Phe Gln Lys Gln His AspHis Leu Thr Arg Gln His Glu Val Gln 100 105 110 Leu Gln Lys His Leu LysGln Gln Gln Glu Met Leu Ala Ala Lys Gln 115 120 125 Gln Gln Glu Met LeuAla Ala Lys Arg Gln Gln Glu Leu Glu Gln Gln 130 135 140 Arg Gln Arg GluGln Gln Arg Gln Glu Glu Leu Glu Lys Gln Arg Leu 145 150 155 160 Glu GlnGln Leu Leu Ile Leu Arg Asn Lys Glu Lys Ser Lys Glu Ser 165 170 175 AlaIle Ala Ser Thr Glu Val Lys Leu Arg Leu Gln Glu Phe Leu Leu 180 185 190Ser Lys Ser Lys Glu Pro Thr Pro Gly Gly Leu Asn His Ser Leu Pro 195 200205 Gln His Pro Lys Cys Trp Gly Ala His His Ala Ser Leu Asp Gln Ser 210215 220 Ser Pro Pro Gln Ser Gly Pro Pro Gly Thr Pro Pro Ser Tyr Lys Leu225 230 235 240 Pro Leu Pro Gly Pro Tyr Asp Ser Arg Asp Asp Phe Pro LeuArg Lys 245 250 255 Thr Ala Ser Glu Pro Asn Leu Lys Val Arg Ser Arg LeuLys Gln Lys 260 265 270 Val Ala Glu Arg Arg Ser Ser Pro Leu Leu Arg ArgLys Asp Gly Thr 275 280 285 Val Ile Ser Thr Phe Lys Lys Arg Ala Val GluIle Thr Gly Ala Gly 290 295 300 Pro Gly Ala Ser Ser Val Cys Asn Ser AlaPro Gly Ser Gly Pro Ser 305 310 315 320 Ser Pro Asn Ser Ser His Ser ThrIle Ala Glu Asn Gly Phe Thr Gly 325 330 335 Ser Val Pro Asn Ile Pro ThrGlu Met Leu Pro Gln His Arg Ala Leu 340 345 350 Pro Leu Asp Ser Ser ProAsn Gln Phe Ser Leu Tyr Thr Ser Pro Ser 355 360 365 Leu Pro Asn Ile SerLeu Gly Leu Gln Ala Thr Val Thr Val Thr Asn 370 375 380 Ser His Leu ThrAla Ser Pro Lys Leu Ser Thr Gln Gln Glu Ala Glu 385 390 395 400 Arg GlnAla Leu Gln Ser Leu Arg Gln Gly Gly Thr Leu Thr Gly Lys 405 410 415 PheMet Ser Thr Ser Ser Ile Pro Gly Cys Leu Leu Gly Val Ala Leu 420 425 430Glu Gly Asp Gly Ser Pro His Gly His Ala Ser Leu Leu Gln His Val 435 440445 Leu Leu Leu Glu Gln Ala Arg Gln Gln Ser Thr Leu Ile Ala Val Pro 450455 460 Leu His Gly Gln Ser Pro Leu Val Thr Gly Glu Arg Val Ala Thr Ser465 470 475 480 Met Arg Thr Val Gly Lys Leu Pro Arg His Arg Pro Leu SerArg Thr 485 490 495 Gln Ser Ser Pro Leu Pro Gln Ser Pro Gln Ala Leu GlnGln Leu Val 500 505 510 Met Gln Gln Gln His Gln Gln Phe Leu Glu Lys GlnLys Gln Gln Gln 515 520 525 Leu Gln Leu Gly Lys Ile Leu Thr Lys Thr GlyGlu Leu Pro Arg Gln 530 535 540 Pro Thr Thr His Pro Glu Glu Thr Glu GluGlu Leu Thr Glu Gln Gln 545 550 555 560 Glu Val Leu Leu Gly Glu Gly AlaLeu Thr Met Pro Arg Glu Gly Ser 565 570 575 Thr Glu Ser Glu Ser Thr GlnGlu Asp Leu Glu Glu Glu Asp Glu Glu 580 585 590 Asp Asp Gly Glu Glu GluGlu Asp Cys Ile Gln Val Lys Asp Glu Glu 595 600 605 Gly Glu Ser Gly AlaGlu Glu Gly Pro Asp Leu Glu Glu Pro Gly Ala 610 615 620 Gly Tyr Lys LysLeu Phe Ser Asp Ala Gln Pro Leu Gln Pro Leu Gln 625 630 635 640 Val TyrGln Ala Pro Leu Ser Leu Ala Thr Val Pro His Gln Ala Leu 645 650 655 GlyArg Thr Gln Ser Ser Pro Ala Ala Pro Gly Gly Met Lys Ser Pro 660 665 670Pro Asp Gln Pro Val Lys His Leu Phe Thr Thr Gly Pro Ile Ser Gln 675 680685 Lys Met Tyr Ala Val Leu Pro Cys Gly Gly Ile Gly Val Asp Ser Asp 690695 700 Thr Val Trp Asn Glu Met His Ser Ser Ser Ala Val Arg Met Ala Val705 710 715 720 Gly Cys Leu Leu Glu Leu Ala Phe Lys Val Ala Ala Gly GluLeu Lys 725 730 735 Asn Gly Phe Ala Ile Ile Arg Pro Pro Gly His His AlaGlu Glu Ser 740 745 750 Thr Ala Met Gly Phe Cys Phe Phe Asn Ser Val AlaIle Thr Ala Lys 755 760 765 Leu Leu Gln Gln Lys Leu Asn Val Gly Lys ValLeu Ile Val Asp Trp 770 775 780 Asp Ile His His Gly Asn Gly Thr Gln GlnAla Phe Tyr Asn Asp Pro 785 790 795 800 Ser Val Leu Tyr Ile Ser Leu HisArg Tyr Asp Asn Gly Asn Phe Phe 805 810 815 Pro Gly Ser Gly Ala Pro GluGlu Val Gly Gly Gly Pro Gly Val Gly 820 825 830 Tyr Asn Val Asn Val AlaTrp Thr Gly Gly Val Asp Pro Pro Ile Gly 835 840 845 Asp Val Glu Tyr LeuThr Ala Phe Arg Thr Val Val Met Pro Ile Ala 850 855 860 His Glu Phe SerPro Asp Val Val Leu Val Ser Ala Gly Phe Asp Ala 865 870 875 880 Val GluGly His Leu Ser Pro Leu Gly Gly Tyr Ser Val Thr Ala Arg 885 890 895 CysPhe Gly His Leu Thr Arg Gln Leu Met Thr Leu Ala Gly Gly Arg 900 905 910Val Val Leu Ala Leu Glu Gly Gly His Asp Leu Thr Ala Ile Cys Asp 915 920925 Ala Ser Glu Ala Cys Val Ser Ala Leu Leu Ser Val Glu Leu Gln Pro 930935 940 Leu Asp Glu Ala Val Leu Gln Gln Lys Pro Asn Ile Asn Ala Val Ala945 950 955 960 Thr Leu Glu Lys Val Ile Glu Ile Gln Ser Lys His Trp SerCys Val 965 970 975 Gln Lys Phe Ala Ala Gly Leu Gly Arg Ser Leu Arg GluAla Gln Ala 980 985 990 Gly Glu Thr Glu Glu Ala Glu Thr Val Ser Ala MetAla Leu Leu Ser 995 1000 1005 Val Gly Ala Glu Gln Ala Gln Ala Ala AlaAla Arg Glu His Ser Pro 1010 1015 1020 Arg Pro Ala Glu Glu Pro Met GluGln Glu Pro Ala Leu 1025 1030 1035 6 1215 PRT Homo sapiens 6 Met Thr SerThr Gly Gln Asp Ser Thr Thr Thr Arg Gln Arg Arg Ser 1 5 10 15 Arg GlnAsn Pro Gln Ser Pro Pro Gln Asp Ser Ser Val Thr Ser Lys 20 25 30 Arg AsnIle Lys Lys Gly Ala Val Pro Arg Ser Ile Pro Asn Leu Ala 35 40 45 Glu ValLys Lys Lys Gly Lys Met Lys Lys Leu Gly Gln Ala Met Glu 50 55 60 Glu AspLeu Ile Val Gly Leu Gln Gly Met Asp Leu Asn Leu Glu Ala 65 70 75 80 GluAla Leu Ala Gly Thr Gly Leu Val Leu Asp Glu Gln Leu Asn Glu 85 90 95 PheHis Cys Leu Trp Asp Asp Ser Phe Pro Glu Gly Pro Glu Arg Leu 100 105 110His Ala Ile Lys Glu Gln Leu Ile Gln Glu Gly Leu Leu Asp Arg Cys 115 120125 Val Ser Phe Gln Ala Arg Phe Ala Glu Lys Glu Glu Leu Met Leu Val 130135 140 His Ser Leu Glu Tyr Ile Asp Leu Met Glu Thr Thr Gln Tyr Met Asn145 150 155 160 Glu Gly Glu Leu Arg Val Leu Ala Asp Thr Tyr Asp Ser ValTyr Leu 165 170 175 His Pro Asn Ser Tyr Ser Cys Ala Cys Leu Ala Ser GlySer Val Leu 180 185 190 Arg Leu Val Asp Ala Val Leu Gly Ala Glu Ile ArgAsn Gly Met Ala 195 200 205 Ile Ile Arg Pro Pro Gly His His Ala Gln HisSer Leu Met Asp Gly 210 215 220 Tyr Cys Met Phe Asn His Val Ala Val AlaAla Arg Tyr Ala Gln Gln 225 230 235 240 Lys His Arg Ile Arg Arg Val LeuIle Val Asp Trp Asp Val His His 245 250 255 Gly Gln Gly Thr Gln Phe ThrPhe Asp Gln Asp Pro Ser Val Leu Tyr 260 265 270 Phe Ser Ile His Arg TyrGlu Gln Gly Arg Phe Trp Pro His Leu Lys 275 280 285 Ala Ser Asn Trp SerThr Thr Gly Phe Gly Gln Gly Gln Gly Tyr Thr 290 295 300 Ile Asn Val ProTrp Asn Gln Val Gly Met Arg Asp Ala Asp Tyr Ile 305 310 315 320 Ala AlaPhe Leu His Val Leu Leu Pro Val Ala Leu Glu Phe Gln Pro 325 330 335 GlnLeu Val Leu Val Ala Ala Gly Phe Asp Ala Leu Gln Gly Asp Pro 340 345 350Lys Gly Glu Met Ala Ala Thr Pro Ala Gly Phe Ala Gln Leu Thr His 355 360365 Leu Leu Met Gly Leu Ala Gly Gly Lys Leu Ile Leu Ser Leu Glu Gly 370375 380 Gly Tyr Asn Leu Arg Ala Leu Ala Glu Gly Val Ser Ala Ser Leu His385 390 395 400 Thr Leu Leu Gly Asp Pro Cys Pro Met Leu Glu Ser Pro GlyAla Pro 405 410 415 Cys Arg Ser Ala Gln Ala Ser Val Ser Cys Ala Leu GluAla Leu Glu 420 425 430 Pro Phe Trp Glu Val Leu Val Arg Ser Thr Glu ThrVal Glu Arg Asp 435 440 445 Asn Met Glu Glu Asp Asn Val Glu Glu Ser GluGlu Glu Gly Pro Trp 450 455 460 Glu Pro Pro Val Leu Pro Ile Leu Thr TrpPro Val Leu Gln Ser Arg 465 470 475 480 Thr Gly Leu Val Tyr Asp Gln AsnMet Met Asn His Cys Asn Leu Trp 485 490 495 Asp Ser His His Pro Glu ValPro Gln Arg Ile Leu Arg Ile Met Cys 500 505 510 Arg Leu Glu Glu Leu GlyLeu Ala Gly Arg Cys Leu Thr Leu Thr Pro 515 520 525 Arg Pro Ala Thr GluAla Glu Leu Leu Thr Cys His Ser Ala Glu Tyr 530 535 540 Val Gly His LeuArg Ala Thr Glu Lys Met Lys Thr Arg Glu Leu His 545 550 555 560 Arg GluSer Ser Asn Phe Asp Ser Ile Tyr Ile Cys Pro Ser Thr Phe 565 570 575 AlaCys Ala Gln Leu Ala Thr Gly Ala Ala Cys Arg Leu Val Glu Ala 580 585 590Val Leu Ser Gly Glu Val Leu Asn Gly Ala Ala Val Val Arg Pro Pro 595 600605 Gly His His Ala Glu Gln Asp Ala Ala Cys Gly Phe Cys Phe Phe Asn 610615 620 Ser Val Ala Val Ala Ala Arg His Ala Gln Thr Ile Ser Gly His Ala625 630 635 640 Leu Arg Ile Leu Ile Val Asp Trp Asp Val His His Gly AsnGly Thr 645 650 655 Gln His Met Phe Glu Asp Asp Pro Ser Val Leu Tyr ValSer Leu His 660 665 670 Arg Tyr Asp His Gly Thr Phe Phe Pro Met Gly AspGlu Gly Ala Ser 675 680 685 Ser Gln Ile Gly Arg Ala Ala Gly Thr Gly PheThr Val Asn Val Ala 690 695 700 Trp Asn Gly Pro Arg Met Gly Asp Ala AspTyr Leu Ala Ala Trp His 705 710 715 720 Arg Leu Val Leu Pro Ile Ala TyrGlu Phe Asn Pro Glu Leu Val Leu 725 730 735 Val Ser Ala Gly Phe Asp AlaAla Arg Gly Asp Pro Leu Gly Gly Cys 740 745 750 Gln Val Ser Pro Glu GlyTyr Ala His Leu Thr His Leu Leu Met Gly 755 760 765 Leu Ala Ser Gly ArgIle Ile Leu Ile Leu Glu Gly Gly Tyr Asn Leu 770 775 780 Thr Ser Ile SerGlu Ser Met Ala Ala Cys Thr Arg Ser Leu Leu Gly 785 790 795 800 Asp ProPro Pro Leu Leu Thr Leu Pro Arg Pro Pro Leu Ser Gly Ala 805 810 815 LeuAla Ser Ile Thr Glu Thr Ile Gln Val His Arg Arg Tyr Trp Arg 820 825 830Ser Leu Arg Val Met Lys Val Glu Asp Arg Glu Gly Pro Ser Ser Ser 835 840845 Lys Leu Val Thr Lys Lys Ala Pro Gln Pro Ala Lys Pro Arg Leu Ala 850855 860 Glu Arg Met Thr Thr Arg Glu Lys Lys Val Leu Glu Ala Gly Met Gly865 870 875 880 Lys Val Thr Ser Ala Ser Phe Gly Glu Glu Ser Thr Pro GlyGln Thr 885 890 895 Asn Ser Glu Thr Ala Val Val Ala Leu Thr Gln Asp GlnPro Ser Glu 900 905 910 Ala Ala Thr Gly Gly Ala Thr Leu Ala Gln Thr IleSer Glu Ala Ala 915 920 925 Ile Gly Gly Ala Met Leu Gly Gln Thr Thr SerGlu Glu Ala Val Gly 930 935 940 Gly Ala Thr Pro Asp Gln Thr Thr Ser GluGlu Thr Val Gly Gly Ala 945 950 955 960 Ile Leu Asp Gln Thr Thr Ser GluAsp Ala Val Gly Gly Ala Thr Leu 965 970 975 Gly Gln Thr Thr Ser Glu GluAla Val Gly Gly Ala Thr Leu Ala Gln 980 985 990 Thr Thr Ser Glu Ala AlaMet Glu Gly Ala Thr Leu Asp Gln Thr Thr 995 1000 1005 Ser Glu Glu AlaPro Gly Gly Thr Glu Leu Ile Gln Thr Pro Leu Ala 1010 1015 1020 Ser SerThr Asp His Gln Thr Pro Pro Thr Ser Pro Val Gln Gly Thr 1025 1030 10351040 Thr Pro Gln Ile Ser Pro Ser Thr Leu Ile Gly Ser Leu Arg Thr Leu1045 1050 1055 Glu Leu Gly Ser Glu Ser Gln Gly Ala Ser Glu Ser Gln AlaPro Gly 1060 1065 1070 Glu Glu Asn Leu Leu Gly Glu Ala Ala Gly Gly GlnAsp Met Ala Asp 1075 1080 1085 Ser Met Leu Met Gln Gly Ser Arg Gly LeuThr Asp Gln Ala Ile Phe 1090 1095 1100 Tyr Ala Val Thr Pro Leu Pro TrpCys Pro His Leu Val Ala Val Cys 1105 1110 1115 1120 Pro Ile Pro Ala AlaGly Leu Asp Val Thr Gln Pro Cys Gly Asp Cys 1125 1130 1135 Gly Thr IleGln Glu Asn Trp Val Cys Leu Ser Cys Tyr Gln Val Tyr 1140 1145 1150 CysGly Arg Tyr Ile Asn Gly His Met Leu Gln His His Gly Asn Ser 1155 11601165 Gly His Pro Leu Val Leu Ser Tyr Ile Asp Leu Ser Ala Trp Cys Tyr1170 1175 1180 Tyr Cys Gln Ala Tyr Val His His Gln Ala Leu Leu Asp ValLys Asn 1185 1190 1195 1200 Ile Ala His Gln Asn Lys Phe Gly Glu Asp MetPro His Pro His 1205 1210 1215 7 952 PRT Homo sapiens 7 Met Asp Leu ArgVal Gly Gln Arg Pro Pro Val Glu Pro Pro Pro Glu 1 5 10 15 Pro Thr LeuLeu Ala Leu Gln Arg Pro Gln Arg Leu His His His Leu 20 25 30 Phe Leu AlaGly Leu Gln Gln Gln Arg Ser Val Glu Pro Met Arg Leu 35 40 45 Ser Met AspThr Pro Met Pro Glu Leu Gln Val Gly Pro Gln Glu Gln 50 55 60 Glu Leu ArgGln Leu Leu His Lys Asp Lys Ser Lys Arg Ser Ala Val 65 70 75 80 Ala SerSer Val Val Lys Gln Lys Leu Ala Glu Val Ile Leu Lys Lys 85 90 95 Gln GlnAla Ala Leu Glu Arg Thr Val His Pro Asn Ser Pro Gly Ile 100 105 110 ProTyr Arg Thr Leu Glu Pro Leu Glu Thr Glu Gly Ala Thr Arg Ser 115 120 125Met Leu Ser Ser Phe Leu Pro Pro Val Pro Ser Leu Pro Ser Asp Pro 130 135140 Pro Glu His Phe Pro Leu Arg Lys Thr Val Ser Glu Pro Asn Leu Lys 145150 155 160 Leu Arg Tyr Lys Pro Lys Lys Ser Leu Glu Arg Arg Lys Asn ProLeu 165 170 175 Leu Arg Lys Glu Ser Ala Pro Pro Ser Leu Arg Arg Arg ProAla Glu 180 185 190 Thr Leu Gly Asp Ser Ser Pro Ser Ser Ser Ser Thr ProAla Ser Gly 195 200 205 Cys Ser Ser Pro Asn Asp Ser Glu His Gly Pro AsnPro Ile Leu Gly 210 215 220 Ser Glu Ala Leu Leu Gly Gln Arg Leu Arg LeuGln Glu Thr Ser Val 225 230 235 240 Ala Pro Phe Ala Leu Pro Thr Val SerLeu Leu Pro Ala Ile Thr Leu 245 250 255 Gly Leu Pro Ala Pro Ala Arg AlaAsp Ser Asp Arg Arg Thr His Pro 260 265 270 Thr Leu Gly Pro Arg Gly ProIle Leu Gly Ser Pro His Thr Pro Leu 275 280 285 Phe Leu Pro His Gly LeuGlu Pro Glu Ala Gly Gly Thr Leu Pro Ser 290 295 300 Arg Leu Gln Pro IleLeu Leu Leu Asp Pro Ser Gly Ser His Ala Pro 305 310 315 320 Leu Leu ThrVal Pro Gly Leu Gly Pro Leu Pro Phe His Phe Ala Gln 325 330 335 Ser LeuMet Thr Thr Glu Arg Leu Ser Gly Ser Gly Leu His Trp Pro 340 345 350 LeuSer Arg Thr Arg Ser Glu Pro Leu Pro Pro Ser Ala Thr Ala Pro 355 360 365Pro Pro Pro Gly Pro Met Gln Pro Arg Leu Glu Gln Leu Lys Thr His 370 375380 Val Gln Val Ile Lys Arg Ser Ala Lys Pro Ser Glu Lys Pro Arg Leu 385390 395 400 Arg Gln Ile Pro Ser Ala Glu Asp Leu Glu Thr Asp Gly Gly GlyPro 405 410 415 Gly Gln Val Val Asp Asp Gly Leu Glu His Arg Glu Leu GlyHis Gly 420 425 430 Gln Pro Glu Ala Arg Gly Pro Ala Pro Leu Gln Gln HisPro Gln Val 435 440 445 Leu Leu Trp Glu Gln Gln Arg Leu Ala Gly Arg LeuPro Arg Gly Ser 450 455 460 Thr Gly Asp Thr Val Leu Leu Pro Leu Ala GlnGly Gly His Arg Pro 465 470 475 480 Leu Ser Arg Ala Gln Ser Ser Pro AlaAla Pro Ala Ser Leu Ser Ala 485 490 495 Pro Glu Pro Ala Ser Gln Ala ArgVal Leu Ser Ser Ser Glu Thr Pro 500 505 510 Ala Arg Thr Leu Pro Phe ThrThr Gly Leu Ile Tyr Asp Ser Val Met 515 520 525 Leu Lys His Gln Cys SerCys Gly Asp Asn Ser Arg His Pro Glu His 530 535 540 Ala Gly Arg Ile GlnSer Ile Trp Ser Arg Leu Gln Glu Arg Gly Leu 545 550 555 560 Arg Ser GlnCys Glu Cys Leu Arg Gly Arg Lys Ala Ser Leu Glu Glu 565 570 575 Leu GlnSer Val His Ser Glu Arg His Val Leu Leu Tyr Gly Thr Asn 580 585 590 ProLeu Ser Arg Leu Lys Leu Asp Asn Gly Lys Leu Ala Gly Leu Leu 595 600 605Ala Gln Arg Met Phe Val Met Leu Pro Cys Gly Gly Val Gly Val Asp 610 615620 Thr Asp Thr Ile Trp Asn Glu Leu His Ser Ser Asn Ala Ala Arg Trp 625630 635 640 Ala Ala Gly Ser Val Thr Asp Leu Ala Phe Lys Val Ala Ser ArgGlu 645 650 655 Leu Lys Asn Gly Phe Ala Val Val Arg Pro Pro Gly His HisAla Asp 660 665 670 His Ser Thr Ala Met Gly Phe Cys Phe Phe Asn Ser ValAla Ile Ala 675 680 685 Cys Arg Gln Leu Gln Gln Gln Ser Lys Ala Ser LysIle Leu Ile Val 690 695 700 Asp Trp Asp Val His His Gly Asn Gly Thr GlnGln Thr Phe Tyr Gln 705 710 715 720 Asp Pro Ser Val Leu Tyr Ile Ser LeuHis Arg His Asp Asp Gly Asn 725 730 735 Phe Phe Pro Gly Ser Gly Ala ValAsp Glu Val Gly Ala Gly Ser Gly 740 745 750 Glu Gly Phe Asn Val Asn ValAla Trp Ala Gly Gly Leu Asp Pro Pro 755 760 765 Met Gly Asp Pro Glu TyrLeu Ala Ala Phe Arg Ile Val Val Met Pro 770 775 780 Ile Ala Arg Glu PheSer Pro Asp Leu Val Leu Val Ser Ala Gly Phe 785 790 795 800 Asp Ala AlaGlu Gly His Pro Ala Pro Leu Gly Gly Tyr His Val Ser 805 810 815 Ala LysCys Phe Gly Tyr Met Thr Gln Gln Leu Met Asn Leu Ala Gly 820 825 830 GlyAla Val Val Leu Ala Leu Glu Gly Gly His Asp Leu Thr Ala Ile 835 840 845Cys Asp Ala Ser Glu Ala Cys Val Ala Ala Leu Leu Gly Asn Arg Val 850 855860 Asp Pro Leu Ser Glu Glu Gly Trp Lys Gln Lys Pro Asn Leu Asn Ala 865870 875 880 Ile Arg Ser Leu Glu Ala Val Ile Arg Val His Ser Lys Tyr TrpGly 885 890 895 Cys Met Gln Arg Leu Ala Ser Cys Pro Asp Ser Trp Val ProArg Val 900 905 910 Pro Gly Ala Asp Lys Glu Glu Val Glu Ala Val Thr AlaLeu Ala Ser 915 920 925 Leu Ser Val Gly Ile Leu Ala Glu Asp Arg Pro SerGlu Gln Leu Val 930 935 940 Glu Glu Glu Glu Pro Met Asn Leu 945 950 8377 PRT Homo sapiens 8 Met Glu Glu Pro Glu Glu Pro Ala Asp Ser Gly GlnSer Leu Val Pro 1 5 10 15 Val Tyr Ile Tyr Ser Pro Glu Tyr Val Ser MetCys Asp Ser Leu Ala 20 25 30 Lys Ile Pro Lys Arg Ala Ser Met Val His SerLeu Ile Glu Ala Tyr 35 40 45 Ala Leu His Lys Gln Met Arg Ile Val Lys ProLys Val Ala Ser Met 50 55 60 Glu Glu Met Ala Thr Phe His Thr Asp Ala TyrLeu Gln His Leu Gln 65 70 75 80 Lys Val Ser Gln Glu Gly Asp Asp Asp HisPro Asp Ser Ile Glu Tyr 85 90 95 Gly Leu Gly Tyr Asp Cys Pro Ala Thr GluGly Ile Phe Asp Tyr Ala 100 105 110 Ala Ala Ile Gly Gly Ala Thr Ile ThrAla Ala Gln Cys Leu Ile Asp 115 120 125 Gly Met Cys Lys Val Ala Ile AsnTrp Ser Gly Gly Trp His His Ala 130 135 140 Lys Lys Asp Glu Ala Ser GlyPhe Cys Tyr Leu Asn Asp Ala Val Leu 145 150 155 160 Gly Ile Leu Arg LeuArg Arg Lys Phe Glu Arg Ile Leu Tyr Val Asp 165 170 175 Leu Asp Leu HisHis Gly Asp Gly Val Glu Asp Ala Phe Ser Phe Thr 180 185 190 Ser Lys ValMet Thr Val Ser Leu His Lys Phe Ser Pro Gly Phe Phe 195 200 205 Pro GlyThr Gly Asp Val Ser Asp Val Gly Leu Gly Lys Gly Arg Tyr 210 215 220 TyrSer Val Asn Val Pro Ile Gln Asp Gly Ile Gln Asp Glu Lys Tyr 225 230 235240 Tyr Gln Ile Cys Glu Ser Val Leu Lys Glu Val Tyr Gln Ala Phe Asn 245250 255 Pro Lys Ala Val Val Leu Gln Leu Gly Ala Asp Thr Ile Ala Gly Asp260 265 270 Pro Met Cys Ser Phe Asn Met Thr Pro Val Gly Ile Gly Lys CysLeu 275 280 285 Lys Tyr Ile Leu Gln Trp Gln Leu Ala Thr Leu Ile Leu GlyGly Gly 290 295 300 Gly Tyr Asn Leu Ala Asn Thr Ala Arg Cys Trp Thr TyrLeu Thr Gly 305 310 315 320 Val Ile Leu Gly Lys Thr Leu Ser Ser Glu IlePro Asp His Glu Phe 325 330 335 Phe Thr Ala Tyr Gly Pro Asp Tyr Val LeuGlu Ile Thr Pro Ser Cys 340 345 350 Arg Pro Asp Arg Asn Glu Pro His ArgIle Gln Gln Ile Leu Asn Tyr 355 360 365 Ile Lys Gly Asn Leu Lys His ValVal 370 375 9 1011 PRT Homo sapeins 9 Met His Ser Met Ile Ser Ser ValAsp Val Lys Ser Glu Val Pro Val 1 5 10 15 Gly Leu Glu Pro Ile Ser ProLeu Asp Leu Arg Thr Asp Leu Arg Met 20 25 30 Met Met Pro Val Val Asp ProVal Val Arg Glu Lys Gln Leu Gln Gln 35 40 45 Glu Leu Leu Leu Ile Gln GlnGln Gln Gln Ile Gln Lys Gln Leu Leu 50 55 60 Ile Ala Glu Phe Gln Lys GlnHis Glu Asn Leu Thr Arg Gln His Gln 65 70 75 80 Ala Gln Leu Gln Glu HisIle Lys Glu Leu Leu Ala Ile Lys Gln Gln 85 90 95 Gln Glu Leu Leu Glu LysGlu Gln Lys Leu Glu Gln Gln Arg Gln Glu 100 105 110 Gln Glu Val Glu ArgHis Arg Arg Glu Gln Gln Leu Pro Pro Leu Arg 115 120 125 Gly Lys Asp ArgGly Arg Glu Arg Ala Val Ala Ser Thr Glu Val Lys 130 135 140 Gln Lys LeuGln Glu Phe Leu Leu Ser Lys Ser Ala Thr Lys Asp Thr 145 150 155 160 ProThr Asn Gly Lys Asn His Ser Val Ser Arg His Pro Lys Leu Trp 165 170 175Tyr Thr Ala Ala His His Thr Ser Leu Asp Gln Ser Ser Pro Pro Leu 180 185190 Ser Gly Thr Ser Pro Ser Tyr Lys Tyr Thr Leu Pro Gly Ala Gln Asp 195200 205 Ala Lys Asp Asp Phe Pro Leu Arg Lys Thr Ala Ser Glu Pro Asn Leu210 215 220 Lys Val Arg Ser Arg Leu Lys Gln Lys Val Ala Glu Arg Arg SerSer 225 230 235 240 Pro Leu Leu Arg Arg Lys Asp Gly Asn Val Val Thr SerPhe Lys Lys 245 250 255 Arg Met Phe Glu Val Thr Glu Ser Ser Val Ser SerSer Ser Pro Gly 260 265 270 Ser Gly Pro Ser Ser Pro Asn Asn Gly Pro ThrGly Ser Val Thr Glu 275 280 285 Asn Glu Thr Ser Val Leu Pro Pro Thr ProHis Ala Glu Gln Met Val 290 295 300 Ser Gln Gln Arg Ile Leu Ile His GluAsp Ser Met Asn Leu Leu Ser 305 310 315 320 Leu Tyr Thr Ser Pro Ser LeuPro Asn Ile Thr Leu Gly Leu Pro Ala 325 330 335 Val Pro Ser Gln Leu AsnAla Ser Asn Ser Leu Lys Glu Lys Gln Lys 340 345 350 Cys Glu Thr Gln ThrLeu Arg Gln Gly Val Pro Leu Pro Gly Gln Tyr 355 360 365 Gly Gly Ser IlePro Ala Ser Ser Ser His Pro His Val Thr Leu Glu 370 375 380 Gly Lys ProPro Asn Ser Ser His Gln Ala Leu Leu Gln His Leu Leu 385 390 395 400 LeuLys Glu Gln Met Arg Gln Gln Lys Leu Leu Val Ala Gly Gly Val 405 410 415Pro Leu His Pro Gln Ser Pro Leu Ala Thr Lys Glu Arg Ile Ser Pro 420 425430 Gly Ile Arg Gly Thr His Lys Leu Pro Arg His Arg Pro Leu Asn Arg 435440 445 Thr Gln Ser Ala Pro Leu Pro Gln Ser Thr Leu Ala Gln Leu Val Ile450 455 460 Gln Gln Gln His Gln Gln Phe Leu Glu Lys Gln Lys Gln Tyr GlnGln 465 470 475 480 Gln Ile His Met Asn Lys Leu Leu Ser Lys Ser Ile GluGln Leu Lys 485 490 495 Gln Pro Gly Ser His Leu Glu Glu Ala Glu Glu GluLeu Gln Gly Asp 500 505 510 Gln Ala Met Gln Glu Asp Arg Ala Pro Ser SerGly Asn Ser Thr Arg 515 520 525 Ser Asp Ser Ser Ala Cys Val Asp Asp ThrLeu Gly Gln Val Gly Ala 530 535 540 Val Lys Val Lys Glu Glu Pro Val AspSer Asp Glu Asp Ala Gln Ile 545 550 555 560 Gln Glu Met Glu Ser Gly GluGln Ala Ala Phe Met Gln Gln Pro Phe 565 570 575 Leu Glu Pro Thr His ThrArg Ala Leu Ser Val Arg Gln Ala Pro Leu 580 585 590 Ala Ala Val Gly MetAsp Gly Leu Glu Lys His Arg Leu Val Ser Arg 595 600 605 Thr His Ser SerPro Ala Ala Ser Val Leu Pro His Pro Ala Met Asp 610 615 620 Arg Pro LeuGln Pro Gly Ser Ala Thr Gly Ile Ala Tyr Asp Pro Leu 625 630 635 640 MetLeu Lys His Gln Cys Val Cys Gly Asn Ser Thr Thr His Pro Glu 645 650 655His Ala Gly Arg Ile Gln Ser Ile Trp Ser Arg Leu Gln Glu Thr Gly 660 665670 Leu Leu Asn Lys Cys Glu Arg Ile Gln Gly Arg Lys Ala Ser Leu Glu 675680 685 Glu Ile Gln Leu Val His Ser Glu His His Ser Leu Leu Tyr Gly Thr690 695 700 Asn Pro Leu Asp Gly Gln Lys Leu Asp Pro Arg Ile Leu Leu GlyAsp 705 710 715 720 Asp Ser Gln Lys Phe Phe Ser Ser Leu Pro Cys Gly GlyLeu Gly Val 725 730 735 Asp Ser Asp Thr Ile Trp Asn Glu Leu His Ser SerGly Ala Ala Arg 740 745 750 Met Ala Val Gly Cys Val Ile Glu Leu Ala SerLys Val Ala Ser Gly 755 760 765 Glu Leu Lys Asn Gly Phe Ala Val Val ArgPro Pro Gly His His Ala 770 775 780 Glu Glu Ser Thr Ala Met Gly Phe CysPhe Phe Asn Ser Val Ala Ile 785 790 795 800 Thr Ala Lys Tyr Leu Arg AspGln Leu Asn Ile Ser Lys Ile Leu Ile 805 810 815 Val Asp Leu Asp Val HisHis Gly Asn Gly Thr Gln Gln Ala Phe Tyr 820 825 830 Ala Asp Pro Ser IleLeu Tyr Ile Ser Leu His Arg Tyr Asp Glu Gly 835 840 845 Asn Phe Phe ProGly Ser Gly Ala Pro Asn Glu Val Gly Thr Gly Leu 850 855 860 Gly Glu GlyTyr Asn Ile Asn Ile Ala Trp Thr Gly Gly Leu Asp Pro 865 870 875 880 ProMet Gly Asp Val Glu Tyr Leu Glu Ala Phe Arg Thr Ile Val Lys 885 890 895Pro Val Ala Lys Glu Phe Asp Pro Asp Met Val Leu Val Ser Ala Gly 900 905910 Phe Asp Ala Leu Glu Gly His Thr Pro Pro Leu Gly Gly Tyr Lys Val 915920 925 Thr Ala Lys Cys Phe Gly His Leu Thr Lys Gln Leu Met Thr Leu Ala930 935 940 Asp Gly Arg Val Val Leu Ala Leu Glu Gly Gly His Asp Leu ThrAla 945 950 955 960 Ile Cys Asp Ala Ser Glu Ala Cys Val Asn Ala Leu LeuGly Asn Glu 965 970 975 Leu Glu Pro Leu Ala Glu Asp Ile Leu His Gln SerPro Asn Met Asn 980 985 990 Ala Val Ile Ser Leu Gln Lys Ile Ile Glu IleGln Ser Met Ser Leu 995 1000 1005 Lys Phe Ser 1010 10 669 PRT Homosapiens 10 Met Gly Thr Ala Leu Val Tyr His Glu Asp Met Thr Ala Thr ArgLeu 1 5 10 15 Leu Trp Asp Asp Pro Glu Cys Glu Ile Glu Arg Pro Glu ArgLeu Thr 20 25 30 Ala Ala Leu Asp Arg Leu Arg Gln Arg Gly Leu Glu Gln ArgCys Leu 35 40 45 Arg Leu Ser Ala Arg Glu Ala Ser Glu Glu Glu Leu Gly LeuVal His 50 55 60 Ser Pro Glu Tyr Val Ser Leu Val Arg Glu Thr Gln Val LeuGly Lys 65 70 75 80 Glu Glu Leu Gln Ala Leu Ser Gly Gln Phe Asp Ala IleTyr Phe His 85 90 95 Pro Ser Thr Phe His Cys Ala Arg Leu Ala Ala Gly AlaGly Leu Gln 100 105 110 Leu Val Asp Ala Val Leu Thr Gly Ala Val Gln AsnGly Leu Ala Leu 115 120 125 Val Arg Pro Pro Gly His His Gly Gln Arg AlaAla Ala Asn Gly Phe 130 135 140 Cys Val Phe Asn Asn Val Ala Ile Ala AlaAla His Ala Lys Gln Lys 145 150 155 160 His Gly Leu His Arg Ile Leu ValVal Asp Trp Asp Val His His Gly 165 170 175 Gln Gly Ile Gln Tyr Leu PheGlu Asp Asp Pro Ser Val Leu Tyr Phe 180 185 190 Ser Trp His Arg Tyr GluHis Gly Arg Phe Trp Pro Phe Leu Arg Glu 195 200 205 Ser Asp Ala Asp AlaVal Gly Arg Gly Gln Gly Leu Gly Phe Thr Val 210 215 220 Asn Leu Pro TrpAsn Gln Val Gly Met Gly Asn Ala Asp Tyr Val Ala 225 230 235 240 Ala PheLeu His Leu Leu Leu Pro Leu Ala Phe Glu Phe Asp Pro Glu 245 250 255 LeuVal Leu Val Ser Ala Gly Phe Asp Ser Ala Ile Gly Asp Pro Glu 260 265 270Gly Gln Met Gln Ala Thr Pro Glu Cys Phe Ala His Leu Thr Gln Leu 275 280285 Leu Gln Val Leu Ala Gly Gly Arg Val Cys Ala Val Leu Glu Gly Gly 290295 300 Tyr His Leu Glu Ser Leu Ala Glu Ser Val Cys Met Thr Val Gln Thr305 310 315 320 Leu Leu Gly Asp Pro Ala Pro Pro Leu Ser Gly Pro Met AlaPro Cys 325 330 335 Gln Ser Ala Leu Glu Ser Ile Gln Ser Ala Arg Ala AlaGln Ala Pro 340 345 350 His Trp Lys Ser Leu Gln Gln Gln Asp Val Thr AlaVal Pro Met Ser 355 360 365 Pro Ser Ser His Ser Pro Glu Gly Arg Pro ProPro Leu Leu Pro Gly 370 375 380 Gly Pro Val Cys Lys Ala Ala Ala Ser AlaPro Ser Ser Leu Leu Asp 385 390 395 400 Gln Pro Cys Leu Cys Pro Ala ProSer Val Arg Thr Ala Val Ala Leu 405 410 415 Thr Thr Pro Asp Ile Thr LeuVal Leu Pro Pro Asp Val Ile Gln Gln 420 425 430 Glu Ala Ser Ala Leu ArgGlu Glu Thr Glu Ala Trp Ala Arg Pro His 435 440 445 Glu Ser Leu Ala ArgGlu Glu Ala Leu Thr Ala Leu Gly Lys Leu Leu 450 455 460 Tyr Leu Leu AspGly Met Leu Asp Gly Gln Val Asn Ser Gly Ile Ala 465 470 475 480 Ala ThrPro Ala Ser Ala Ala Ala Ala Thr Leu Asp Val Ala Val Arg 485 490 495 ArgGly Leu Ser His Gly Ala Gln Arg Leu Leu Cys Val Ala Leu Gly 500 505 510Gln Leu Asp Arg Pro Pro Asp Leu Ala His Asp Gly Arg Ser Leu Trp 515 520525 Leu Asn Ile Arg Gly Lys Glu Ala Ala Ala Leu Ser Met Phe His Val 530535 540 Ser Thr Pro Leu Pro Val Met Thr Gly Gly Phe Leu Ser Cys Ile Leu545 550 555 560 Gly Leu Val Leu Pro Leu Ala Tyr Gly Phe Gln Pro Asp LeuVal Leu 565 570 575 Val Ala Leu Gly Pro Gly His Gly Leu Gln Gly Pro HisAla Ala Leu 580 585 590 Leu Ala Ala Met Leu Arg Gly Leu Ala Gly Gly ArgVal Leu Ala Leu 595 600 605 Leu Glu Glu Asn Ser Thr Pro Gln Leu Ala GlyIle Leu Ala Arg Val 610 615 620 Leu Asn Gly Glu Ala Pro Pro Ser Leu GlyPro Ser Ser Val Ala Ser 625 630 635 640 Pro Glu Asp Val Gln Ala Leu MetTyr Leu Arg Gly Gln Leu Glu Pro 645 650 655 Gln Trp Lys Met Leu Gln CysHis Pro His Leu Val Ala 660 665 11 347 PRT Homo sapiens 11 Met Leu HisThr Thr Gln Leu Tyr Gln His Val Pro Glu Thr Pro Trp 1 5 10 15 Pro IleVal Tyr Ser Pro Arg Tyr Asn Ile Thr Phe Met Gly Leu Glu 20 25 30 Lys LeuHis Pro Phe Asp Ala Gly Lys Trp Gly Lys Val Ile Asn Phe 35 40 45 Leu LysGlu Glu Lys Leu Leu Ser Asp Ser Met Leu Val Glu Ala Arg 50 55 60 Glu AlaSer Glu Glu Asp Leu Leu Val Val His Thr Arg Arg Tyr Leu 65 70 75 80 AsnGlu Leu Lys Trp Ser Phe Ala Val Ala Thr Ile Thr Glu Ile Pro 85 90 95 ProVal Ile Phe Leu Pro Asn Phe Leu Val Gln Arg Lys Val Leu Arg 100 105 110Pro Leu Arg Thr Gln Thr Gly Gly Thr Ile Met Ala Gly Lys Leu Ala 115 120125 Val Glu Arg Gly Trp Ala Ile Asn Val Gly Gly Gly Phe His His Cys 130135 140 Ser Ser Asp Arg Gly Gly Gly Phe Cys Ala Tyr Ala Asp Ile Thr Leu145 150 155 160 Ala Ile Lys Phe Leu Phe Glu Arg Val Glu Gly Ile Ser ArgAla Thr 165 170 175 Ile Ile Asp Leu Asp Ala His Gln Gly Asn Gly His GluArg Asp Phe 180 185 190 Met Asp Asp Lys Arg Val Tyr Ile Met Asp Val TyrAsn Arg His Ile 195 200 205 Tyr Pro Gly Asp Arg Phe Ala Lys Gln Ala IleArg Arg Lys Val Glu 210 215 220 Leu Glu Trp Gly Thr Glu Asp Asp Glu TyrLeu Asp Lys Val Glu Arg 225 230 235 240 Asn Ile Lys Lys Ser Leu Gln GluHis Leu Pro Asp Val Val Val Tyr 245 250 255 Asn Ala Gly Thr Asp Ile LeuGlu Gly Asp Arg Leu Gly Gly Leu Ser 260 265 270 Ile Ser Pro Ala Gly IleVal Lys Arg Asp Glu Leu Val Phe Arg Met 275 280 285 Val Arg Gly Arg ArgVal Pro Ile Leu Met Val Thr Ser Gly Gly Tyr 290 295 300 Gln Lys Arg ThrAla Arg Ile Ile Ala Asp Ser Ile Leu Asn Leu Phe 305 310 315 320 Gly LeuGly Leu Ile Gly Pro Glu Ser Pro Ser Val Ser Ala Gln Asn 325 330 335 SerAsp Thr Pro Leu Leu Pro Pro Ala Val Pro 340 345 12 747 PRT Homo sapiens12 Met Ala Asp Glu Ala Ala Leu Ala Leu Gln Pro Gly Gly Ser Pro Ser 1 510 15 Ala Ala Gly Ala Asp Arg Glu Ala Ala Ser Ser Pro Ala Gly Glu Pro 2025 30 Leu Arg Lys Arg Pro Arg Arg Asp Gly Pro Gly Leu Glu Arg Ser Pro 3540 45 Gly Glu Pro Gly Gly Ala Ala Pro Glu Arg Glu Val Pro Ala Ala Ala 5055 60 Arg Gly Cys Pro Gly Ala Ala Ala Ala Ala Leu Trp Arg Glu Ala Glu 6570 75 80 Ala Glu Ala Ala Ala Ala Gly Gly Glu Gln Glu Ala Gln Ala Thr Ala85 90 95 Ala Ala Gly Glu Gly Asp Asn Gly Pro Gly Leu Gln Gly Pro Ser Arg100 105 110 Glu Pro Pro Leu Ala Asp Asn Leu Tyr Asp Glu Asp Asp Asp AspGlu 115 120 125 Gly Glu Glu Glu Glu Glu Ala Ala Ala Ala Ala Ile Gly TyrArg Asp 130 135 140 Asn Leu Leu Phe Gly Asp Glu Ile Ile Thr Asn Gly PheHis Ser Cys 145 150 155 160 Glu Ser Asp Glu Glu Asp Arg Ala Ser His AlaSer Ser Ser Asp Trp 165 170 175 Thr Pro Arg Pro Arg Ile Gly Pro Tyr ThrPhe Val Gln Gln His Leu 180 185 190 Met Ile Gly Thr Asp Pro Arg Thr IleLeu Lys Asp Leu Leu Pro Glu 195 200 205 Thr Ile Pro Pro Pro Glu Leu AspAsp Met Thr Leu Trp Gln Ile Val 210 215 220 Ile Asn Ile Leu Ser Glu ProPro Lys Arg Lys Lys Arg Lys Asp Ile 225 230 235 240 Asn Thr Ile Glu AspAla Val Lys Leu Leu Gln Glu Cys Lys Lys Ile 245 250 255 Ile Val Leu ThrGly Ala Gly Val Ser Val Ser Cys Gly Ile Pro Asp 260 265 270 Phe Arg SerArg Asp Gly Ile Tyr Ala Arg Leu Ala Val Asp Phe Pro 275 280 285 Asp LeuPro Asp Pro Gln Ala Met Phe Asp Ile Glu Tyr Phe Arg Lys 290 295 300 AspPro Arg Pro Phe Phe Lys Phe Ala Lys Glu Ile Tyr Pro Gly Gln 305 310 315320 Phe Gln Pro Ser Leu Cys His Lys Phe Ile Ala Leu Ser Asp Lys Glu 325330 335 Gly Lys Leu Leu Arg Asn Tyr Thr Gln Asn Ile Asp Thr Leu Glu Gln340 345 350 Val Ala Gly Ile Gln Arg Ile Ile Gln Cys His Gly Ser Phe AlaThr 355 360 365 Ala Ser Cys Leu Ile Cys Lys Tyr Lys Val Asp Cys Glu AlaVal Arg 370 375 380 Gly Asp Ile Phe Asn Gln Val Val Pro Arg Cys Pro ArgCys Pro Ala 385 390 395 400 Asp Glu Pro Leu Ala Ile Met Lys Pro Glu IleVal Phe Phe Gly Glu 405 410 415 Asn Leu Pro Glu Gln Phe His Arg Ala MetLys Tyr Asp Lys Asp Glu 420 425 430 Val Asp Leu Leu Ile Val Ile Gly SerSer Leu Lys Val Arg Pro Val 435 440 445 Ala Leu Ile Pro Ser Ser Ile ProHis Glu Val Pro Gln Ile Leu Ile 450 455 460 Asn Arg Glu Pro Leu Pro HisLeu His Phe Asp Val Glu Leu Leu Gly 465 470 475 480 Asp Cys Asp Val IleIle Asn Glu Leu Cys His Arg Leu Gly Gly Glu 485 490 495 Tyr Ala Lys LeuCys Cys Asn Pro Val Lys Leu Ser Glu Ile Thr Glu 500 505 510 Lys Pro ProArg Thr Gln Lys Glu Leu Ala Tyr Leu Ser Glu Leu Pro 515 520 525 Pro ThrPro Leu His Val Ser Glu Asp Ser Ser Ser Pro Glu Arg Thr 530 535 540 SerPro Pro Asp Ser Ser Val Ile Val Thr Leu Leu Asp Gln Ala Ala 545 550 555560 Lys Ser Asn Asp Asp Leu Asp Val Ser Glu Ser Lys Gly Cys Met Glu 565570 575 Glu Lys Pro Gln Glu Val Gln Thr Ser Arg Asn Val Glu Ser Ile Ala580 585 590 Glu Gln Met Glu Asn Pro Asp Leu Lys Asn Val Gly Ser Ser ThrGly 595 600 605 Glu Lys Asn Glu Arg Thr Ser Val Ala Gly Thr Val Arg LysCys Trp 610 615 620 Pro Asn Arg Val Ala Lys Glu Gln Ile Ser Arg Arg LeuAsp Gly Asn 625 630 635 640 Gln Tyr Leu Phe Leu Pro Pro Asn Arg Tyr IlePhe His Gly Ala Glu 645 650 655 Val Tyr Ser Asp Ser Glu Asp Asp Val LeuSer Ser Ser Ser Cys Gly 660 665 670 Ser Asn Ser Asp Ser Gly Thr Cys GlnSer Pro Ser Leu Glu Glu Pro 675 680 685 Met Glu Asp Glu Ser Glu Ile GluGlu Phe Tyr Asn Gly Leu Glu Asp 690 695 700 Glu Pro Asp Val Pro Glu ArgAla Gly Gly Ala Gly Phe Gly Thr Asp 705 710 715 720 Gly Asp Asp Gln GluAla Ile Asn Glu Ala Ile Ser Val Lys Gln Glu 725 730 735 Val Thr Asp MetAsn Tyr Pro Ser Asn Lys Ser 740 745 13 389 PRT Homo sapiens 13 Met AlaGlu Pro Asp Pro Ser His Pro Leu Glu Thr Gln Ala Gly Lys 1 5 10 15 ValGln Glu Ala Gln Asp Ser Asp Ser Asp Ser Glu Gly Gly Ala Ala 20 25 30 GlyGly Glu Ala Asp Met Asp Phe Leu Arg Asn Leu Phe Ser Gln Thr 35 40 45 LeuSer Leu Gly Ser Gln Lys Glu Arg Leu Leu Asp Glu Leu Thr Leu 50 55 60 GluGly Val Ala Arg Tyr Met Gln Ser Glu Arg Cys Arg Arg Val Ile 65 70 75 80Cys Leu Val Gly Ala Gly Ile Ser Thr Ser Ala Gly Ile Pro Asp Phe 85 90 95Arg Ser Pro Ser Thr Gly Leu Tyr Asp Asn Leu Glu Lys Tyr His Leu 100 105110 Pro Tyr Pro Glu Ala Ile Phe Glu Ile Ser Tyr Phe Lys Lys His Pro 115120 125 Glu Pro Phe Phe Ala Leu Ala Lys Glu Leu Tyr Pro Gly Gln Phe Lys130 135 140 Pro Thr Ile Cys His Tyr Phe Met Arg Leu Leu Lys Asp Lys GlyLeu 145 150 155 160 Leu Leu Arg Cys Tyr Thr Gln Asn Ile Asp Thr Leu GluArg Ile Ala 165 170 175 Gly Leu Glu Gln Glu Asp Leu Val Glu Ala His GlyThr Phe Tyr Thr 180 185 190 Ser His Cys Val Ser Ala Ser Cys Arg His GluTyr Pro Leu Ser Trp 195 200 205 Met Lys Glu Lys Ile Phe Ser Glu Val ThrPro Lys Cys Glu Asp Cys 210 215 220 Gln Ser Leu Val Lys Pro Asp Ile ValPhe Phe Gly Glu Ser Leu Pro 225 230 235 240 Ala Arg Phe Phe Ser Cys MetGln Ser Asp Phe Leu Lys Val Asp Leu 245 250 255 Leu Leu Val Met Gly ThrSer Leu Gln Val Gln Pro Phe Ala Ser Leu 260 265 270 Ile Ser Lys Ala ProLeu Ser Thr Pro Arg Leu Leu Ile Asn Lys Glu 275 280 285 Lys Ala Gly GlnSer Asp Pro Phe Leu Gly Met Ile Met Gly Leu Gly 290 295 300 Gly Gly MetAsp Phe Asp Ser Lys Lys Ala Tyr Arg Asp Val Ala Trp 305 310 315 320 LeuGly Glu Cys Asp Gln Gly Cys Leu Ala Leu Ala Glu Leu Leu Gly 325 330 335Trp Lys Lys Glu Leu Glu Asp Leu Val Arg Arg Glu His Ala Ser Ile 340 345350 Asp Ala Gln Ser Gly Ala Gly Val Pro Asn Pro Ser Thr Ser Ala Ser 355360 365 Pro Lys Lys Ser Pro Pro Pro Ala Lys Asp Glu Ala Arg Thr Thr Glu370 375 380 Arg Glu Lys Pro Gln 385 14 399 PRT Homo sapiens 14 Met AlaPhe Trp Gly Trp Arg Ala Ala Ala Ala Leu Arg Leu Trp Gly 1 5 10 15 ArgVal Val Glu Arg Val Glu Ala Gly Gly Gly Val Gly Pro Phe Gln 20 25 30 AlaCys Gly Cys Arg Leu Val Leu Gly Gly Arg Asp Asp Val Ser Ala 35 40 45 GlyLeu Arg Gly Ser His Gly Ala Arg Gly Glu Pro Leu Asp Pro Ala 50 55 60 ArgPro Leu Gln Arg Pro Pro Arg Pro Glu Val Pro Arg Ala Phe Arg 65 70 75 80Arg Gln Pro Arg Ala Ala Ala Pro Ser Phe Phe Phe Ser Ser Ile Lys 85 90 95Gly Gly Arg Arg Ser Ile Ser Phe Ser Val Gly Ala Ser Ser Val Val 100 105110 Gly Ser Gly Gly Ser Ser Asp Lys Gly Lys Leu Ser Leu Gln Asp Val 115120 125 Ala Glu Leu Ile Arg Ala Arg Ala Cys Gln Arg Val Val Val Met Val130 135 140 Gly Ala Gly Ile Ser Thr Pro Ser Gly Ile Pro Asp Phe Arg SerPro 145 150 155 160 Gly Ser Gly Leu Tyr Ser Asn Leu Gln Gln Tyr Asp LeuPro Tyr Pro 165 170 175 Glu Ala Ile Phe Glu Leu Pro Phe Phe Phe His AsnPro Lys Pro Phe 180 185 190 Phe Thr Leu Ala Lys Glu Leu Tyr Pro Gly AsnTyr Lys Pro Asn Val 195 200 205 Thr His Tyr Phe Leu Arg Leu Leu His AspLys Gly Leu Leu Leu Arg 210 215 220 Leu Tyr Thr Gln Asn Ile Asp Gly LeuGlu Arg Val Ser Gly Ile Pro 225 230 235 240 Ala Ser Lys Leu Val Glu AlaHis Gly Thr Phe Ala Ser Ala Thr Cys 245 250 255 Thr Val Cys Gln Arg ProPhe Pro Gly Glu Asp Ile Arg Ala Asp Val 260 265 270 Met Ala Asp Arg ValPro Arg Cys Pro Val Cys Thr Gly Val Val Lys 275 280 285 Pro Asp Ile ValPhe Phe Gly Glu Pro Leu Pro Gln Arg Phe Leu Leu 290 295 300 His Val ValAsp Phe Pro Met Ala Asp Leu Leu Leu Ile Leu Gly Thr 305 310 315 320 SerLeu Glu Val Glu Pro Phe Ala Ser Leu Thr Glu Ala Val Arg Ser 325 330 335Ser Val Pro Arg Leu Leu Ile Asn Arg Asp Leu Val Gly Pro Leu Ala 340 345350 Trp His Pro Arg Ser Arg Asp Val Ala Gln Leu Gly Asp Val Val His 355360 365 Gly Val Glu Ser Leu Val Glu Leu Leu Gly Trp Thr Glu Glu Met Arg370 375 380 Asp Leu Val Gln Arg Glu Thr Gly Lys Leu Asp Gly Pro Asp Lys385 390 395 15 314 PRT Homo sapiens 15 Met Lys Met Ser Phe Ala Leu ThrPhe Arg Ser Ala Lys Gly Arg Trp 1 5 10 15 Ile Ala Asn Pro Ser Gln ProCys Ser Lys Ala Ser Ile Gly Leu Phe 20 25 30 Val Pro Ala Ser Pro Pro LeuAsp Pro Glu Lys Val Lys Glu Leu Gln 35 40 45 Arg Phe Ile Thr Leu Ser LysArg Leu Leu Val Met Thr Gly Ala Gly 50 55 60 Ile Ser Thr Glu Ser Gly IlePro Asp Tyr Arg Ser Glu Lys Val Gly 65 70 75 80 Leu Tyr Ala Arg Thr AspArg Arg Pro Ile Gln His Gly Asp Phe Val 85 90 95 Arg Ser Ala Pro Ile ArgGln Arg Tyr Trp Ala Arg Asn Phe Val Gly 100 105 110 Trp Pro Gln Phe SerSer His Gln Pro Asn Pro Ala His Trp Ala Leu 115 120 125 Ser Thr Trp GluLys Leu Gly Lys Leu Tyr Trp Leu Val Thr Gln Asn 130 135 140 Val Asp AlaLeu His Thr Lys Ala Gly Ser Arg Arg Leu Thr Glu Leu 145 150 155 160 HisGly Cys Met Asp Arg Val Leu Cys Leu Asp Cys Gly Glu Gln Thr 165 170 175Pro Arg Gly Val Leu Gln Glu Arg Phe Gln Val Leu Asn Pro Thr Trp 180 185190 Ser Ala Glu Ala His Gly Leu Ala Pro Asp Gly Asp Val Phe Leu Ser 195200 205 Glu Glu Gln Val Arg Ser Phe Gln Val Pro Thr Cys Val Gln Cys Gly210 215 220 Gly His Leu Lys Pro Asp Val Val Phe Phe Gly Asp Thr Val AsnPro 225 230 235 240 Asp Lys Val Asp Phe Val His Lys Arg Val Lys Glu AlaAsp Ser Leu 245 250 255 Leu Val Val Gly Ser Ser Leu Gln Val Tyr Ser GlyTyr Arg Phe Ile 260 265 270 Leu Thr Ala Trp Glu Lys Lys Leu Pro Ile AlaIle Leu Asn Ile Gly 275 280 285 Pro Thr Arg Ser Asp Asp Leu Ala Cys LeuLys Leu Asn Ser Arg Cys 290 295 300 Gly Glu Leu Leu Pro Leu Ile Asp ProCys 305 310 16 310 PRT Homo sapiens 16 Met Arg Pro Leu Gln Ile Val ProSer Arg Leu Ile Ser Gln Leu Tyr 1 5 10 15 Cys Gly Leu Lys Pro Pro AlaSer Thr Arg Asn Gln Ile Cys Leu Lys 20 25 30 Met Ala Arg Pro Ser Ser SerMet Ala Asp Phe Arg Lys Phe Phe Ala 35 40 45 Lys Ala Lys His Ile Val IleIle Ser Gly Ala Gly Val Ser Ala Glu 50 55 60 Ser Gly Val Pro Thr Phe ArgGly Ala Gly Gly Tyr Trp Arg Lys Trp 65 70 75 80 Gln Ala Gln Asp Leu AlaThr Pro Leu Ala Phe Ala His Asn Pro Ser 85 90 95 Arg Val Trp Glu Phe TyrHis Tyr Arg Arg Glu Val Met Gly Ser Lys 100 105 110 Glu Pro Asn Ala GlyHis Arg Ala Ile Ala Glu Cys Glu Thr Arg Leu 115 120 125 Gly Lys Gln GlyArg Arg Val Val Val Ile Thr Gln Asn Ile Asp Glu 130 135 140 Leu His ArgLys Ala Gly Thr Lys Asn Leu Leu Glu Ile His Gly Ser 145 150 155 160 LeuPhe Lys Thr Arg Cys Thr Ser Cys Gly Val Val Ala Glu Asn Tyr 165 170 175Lys Ser Pro Ile Cys Pro Ala Leu Ser Gly Lys Gly Ala Pro Glu Pro 180 185190 Gly Thr Gln Asp Ala Ser Ile Pro Val Glu Lys Leu Pro Arg Cys Glu 195200 205 Glu Ala Gly Cys Gly Gly Leu Leu Arg Pro His Val Val Trp Phe Gly210 215 220 Glu Asn Leu Asp Pro Ala Ile Leu Glu Glu Val Asp Arg Glu LeuAla 225 230 235 240 His Cys Asp Leu Cys Leu Val Val Gly Thr Ser Ser ValVal Tyr Pro 245 250 255 Ala Ala Met Phe Ala Pro Gln Val Ala Ala Arg GlyVal Pro Val Ala 260 265 270 Glu Phe Asn Thr Glu Thr Thr Pro Ala Thr AsnArg Phe Arg Phe His 275 280 285 Phe Gln Gly Pro Cys Gly Thr Thr Leu ProGlu Ala Leu Ala Cys His 290 295 300 Glu Asn Glu Thr Val Ser 305 310 17355 PRT Homo sapiens 17 Met Ser Val Asn Tyr Ala Ala Gly Leu Ser Pro TyrAla Asp Lys Gly 1 5 10 15 Lys Cys Gly Leu Pro Glu Ile Phe Asp Pro ProGlu Glu Leu Glu Arg 20 25 30 Lys Val Trp Glu Leu Ala Arg Leu Val Trp GlnSer Ser Ser Val Val 35 40 45 Phe His Thr Gly Ala Gly Ile Ser Thr Ala SerGly Ile Pro Asp Phe 50 55 60 Arg Gly Pro His Gly Val Trp Thr Met Glu GluArg Gly Leu Ala Pro 65 70 75 80 Lys Phe Asp Thr Thr Phe Glu Ser Ala ArgPro Thr Gln Thr His Met 85 90 95 Ala Leu Val Gln Leu Glu Arg Val Gly LeuLeu Arg Phe Leu Val Ser 100 105 110 Gln Asn Val Asp Gly Leu His Val ArgSer Gly Phe Pro Arg Asp Lys 115 120 125 Leu Ala Glu Leu His Gly Asn MetPhe Val Glu Glu Cys Ala Lys Cys 130 135 140 Lys Thr Gln Tyr Val Arg AspThr Val Val Gly Thr Met Gly Leu Lys 145 150 155 160 Ala Thr Gly Arg LeuCys Thr Val Ala Lys Ala Arg Gly Leu Arg Ala 165 170 175 Cys Arg Gly GluLeu Arg Asp Thr Ile Leu Asp Trp Glu Asp Ser Leu 180 185 190 Pro Asp ArgAsp Leu Ala Leu Ala Asp Glu Ala Ser Arg Asn Ala Asp 195 200 205 Leu SerIle Thr Leu Gly Thr Ser Leu Gln Ile Arg Pro Ser Gly Asn 210 215 220 LeuPro Leu Ala Thr Lys Arg Arg Gly Gly Arg Leu Val Ile Val Asn 225 230 235240 Leu Gln Pro Thr Lys His Asp Arg His Ala Asp Leu Arg Ile His Gly 245250 255 Tyr Val Asp Glu Val Met Thr Arg Leu Met Glu His Leu Gly Leu Glu260 265 270 Ile Pro Ala Trp Asp Gly Pro Arg Val Leu Glu Arg Ala Leu ProPro 275 280 285 Leu Pro Arg Pro Pro Thr Pro Lys Leu Glu Pro Lys Glu GluSer Pro 290 295 300 Thr Arg Ile Asn Gly Ser Ile Pro Ala Gly Pro Lys GlnGlu Pro Cys 305 310 315 320 Ala Gln His Asn Gly Ser Glu Pro Ala Ser ProLys Arg Glu Arg Pro 325 330 335 Thr Ser Pro Ala Pro His Arg Pro Pro LysArg Val Lys Ala Lys Ala 340 345 350 Val Pro Ser 355 18 400 PRT Homosapiens 18 Met Ala Ala Gly Gly Leu Ser Arg Ser Glu Arg Lys Ala Ala GluArg 1 5 10 15 Val Arg Arg Leu Arg Glu Glu Gln Gln Arg Glu Arg Leu ArgGln Val 20 25 30 Ser Arg Ile Leu Arg Lys Ala Ala Ala Glu Arg Ser Ala GluGlu Gly 35 40 45 Arg Leu Leu Ala Glu Ser Ala Asp Leu Val Thr Glu Leu GlnGly Arg 50 55 60 Ser Arg Arg Arg Glu Gly Leu Lys Arg Arg Gln Glu Glu ValCys Asp 65 70 75 80 Asp Pro Glu Glu Leu Arg Gly Lys Val Arg Glu Leu AlaSer Ala Val 85 90 95 Arg Asn Ala Lys Tyr Leu Val Val Tyr Thr Gly Ala GlyIle Ser Thr 100 105 110 Ala Ala Ser Ile Pro Asp Tyr Arg Gly Pro Asn GlyVal Trp Thr Leu 115 120 125 Leu Gln Lys Gly Arg Ser Val Ser Ala Ala AspLeu Ser Glu Ala Glu 130 135 140 Pro Thr Leu Thr His Met Ser Ile Thr ArgLeu His Glu Gln Lys Leu 145 150 155 160 Val Gln His Val Val Ser Gln AsnCys Asp Gly Leu His Leu Arg Ser 165 170 175 Gly Leu Pro Arg Thr Ala IleSer Glu Leu His Gly Asn Met Tyr Ile 180 185 190 Glu Val Cys Thr Ser CysVal Pro Asn Arg Glu Tyr Val Arg Val Phe 195 200 205 Asp Val Thr Glu ArgThr Ala Leu His Arg His Gln Thr Gly Arg Thr 210 215 220 Cys His Lys CysGly Thr Gln Leu Arg Asp Thr Ile Val His Phe Gly 225 230 235 240 Glu ArgGly Thr Leu Gly Gln Pro Leu Asn Trp Glu Ala Ala Thr Glu 245 250 255 AlaAla Ser Arg Ala Asp Thr Ile Leu Cys Leu Gly Ser Ser Leu Lys 260 265 270Val Leu Lys Lys Tyr Pro Arg Leu Trp Cys Met Thr Lys Pro Pro Ser 275 280285 Arg Arg Pro Lys Leu Tyr Ile Val Asn Leu Gln Trp Thr Pro Lys Asp 290295 300 Asp Trp Ala Ala Leu Lys Leu His Gly Lys Cys Asp Asp Val Met Arg305 310 315 320 Leu Leu Met Ala Glu Leu Gly Leu Glu Ile Pro Ala Tyr SerArg Trp 325 330 335 Gln Asp Pro Ile Phe Ser Leu Ala Thr Pro Leu Arg AlaGly Glu Glu 340 345 350 Gly Ser His Ser Arg Lys Ser Leu Cys Arg Ser ArgGlu Glu Ala Pro 355 360 365 Pro Gly Asp Arg Gly Ala Pro Leu Ser Ser AlaPro Ile Leu Gly Gly 370 375 380 Trp Phe Gly Arg Gly Cys Thr Lys Arg ThrLys Arg Lys Lys Val Thr 385 390 395 400

What is claimed is:
 1. A continuous method for measuring the activity ofan enzyme that catalyzes (a) the addition of an acetyl group to aresidue capable of being acetylated or (b) removal of an acetyl groupfrom an acetylated residue which method comprises incubating said enzymewith: (i) a protease; (ii) a polypeptide comprising: (a) a recognitionsite for the protease; (b) a residue, in which the acetylation state ofthe residue modifies the rate of cleavage of the polypeptide by theprotease; and (c) at least one chemical moiety, attached to thepolypeptide, that possesses an optical signal that changes upon cleavageof the polypeptide; and (iii) measuring the change in the opticalsignal.
 2. A continuous method for measuring the activity of an enzymethat catalyzes the addition of an acetyl group to a lysine residue or anenzyme that catalyzes the removal of acetyl group from N^(ε)-acetylatedlysine residue which method comprises incubating said enzyme with: (i) aprotease; (ii) a polypeptide comprising: (a) a recognition site for theprotease; (b) a lysine or acetyllysine residue, in which the acetylationstate of the residue modifies the rate of cleavage of the polypeptide bythe protease; and (c) at least one chemical moiety, attached to thepolypeptide, that possesses an optical signal that changes upon cleavageof the polypeptide; and (iii) measuring the change in the opticalsignal.
 3. A continuous method for measuring the activity of an enzymethat catalyzes the removal of acetyl group from N^(ε)-acetylated lysineresidue which method comprises incubating said enzyme with: (i) aprotease; (ii) a polypeptide comprising: (a) a recognition site for theprotease; (b) an acetyllysine residue, which residue when acetylatedattenuates the rate of cleavage of the polypeptide by the protease; and(c) at least one chemical moiety, attached to the polypeptide, thatpossesses an optical signal that changes upon cleavage of thepolypeptide; and (iii) measuring the change in the optical signal. 4.The method of claim 3 where the enzyme is HDAC1 (SEQ. ID. NO: 1), HDAC2(SEQ. ID. NO: 2), HDAC3 (SEQ. ID. NO: 3), HDAC4 (SEQ. ID. NO: 4), HDAC5(SEQ. ID. NO: 5), HDAC6 (SEQ. ID. NO: 6), HDAC7 (SEQ. ID. NO: 7), HDAC8(SEQ. ID. NO: 8), HDAC9 (SEQ. ID. NO: 9), HDAC10 (SEQ. ID. NO: 10), orHDAC11 (SEQ. ID. NO: 11), any protein with 95% or greater sequencesimilarity any of the said enzymes, or any fragment of any of theenzymes that retains catalytic deacetylase activity.
 5. The method ofclaim 3 where the enzyme is SIRT1 (SEQ. ID. NO: 12), SIRT2 (SEQ. ID. NO:13), SIRT3 (SEQ. ID. NO: 14), SIRT4 (SEQ. ID. NO: 15), SIRT5 (SEQ. ID.NO: 16), SIRT6 (SEQ. ID. NO: 17), or SIRT7 (SEQ. ID. NO: 18), anyprotein with 95% or greater sequence similarity to any of the saidenzymes, or any fragment of any of the enzymes that retains catalyticdeacetylase activity.
 6. The method of claim 3 where the protease is amember of the trypsin family of proteases.
 7. The method of claim 3where the protease is trypsin or thrombin.
 8. The method of claim 3where the optical signal arises from a fluorescent moiety.
 9. The methodof claim 3 where the optical signal arises from optical absorption. 10.The method of claim 3 where the optical signal arises from afluorescence resonance energy transfer.
 11. The method of claim 7 wherethe polypeptide is acetyl-Gly-Ala-(N^(ε)-acetyllysine)-AMC.
 12. Themethod of claim 10 where the polypeptide is(2-aminobenzoyl)-Gly-Ala-(N^(ε)-acetyllysine)-Ala-Ala-(3-(2,4-dinitrophenyl)-2,3-diaminopropionamide).13. The method of claim 3 where the polypeptide is less than or equal to20 amino acids in length.
 14. The method of claim 3 where thepolypeptide is less than or equal to 8 amino acids in length.
 15. Acontinuous method for measuring the activity of a histone deacetylaseenzyme which method comprises incubating the histone deacetylase enzymewith: (i) trypsin; (ii) acetyl-Gly-Ala-(N^(ε)-acetyllysine)-AMC; and(iii) measuring the increase in fluorescence at 460 nm over time, usingan excitation wavelength of 355 nm.
 16. The method of claim 15 where thehistone deacetylase enzyme is HDAC1.
 17. A continuous method formeasuring the inhibitory properties of a test compound towards an enzymethat catalyzes the addition or removal of acetyl group from N^(ε)-acetylated lysine residue which method comprises incubating said enzymewith: (i) a protease; (ii) a polypeptide comprising: (a) a recognitionsite for the protease; (b) a lysine or acetyllysine residue, in whichthe acetylation state of the lysine residue modifies the rate ofcleavage of the polypeptide by the protease; and (c) at least onechemical moiety, attached to the polypeptide, that possesses an opticalsignal that changes upon cleavage of the polypeptide; in the presence ofthe test compound; (iii) measuring the rate of increase in the opticalsignal wherein the difference between the rate of increase of theoptical signal in the presence of the test compound and the rate ofincrease of the optical signal in the absence of the test compound isindicative of inhibitory properties of the test compound.
 18. Acontinuous method for measuring the inhibitory properties of a testcompound towards a histone deacetylase enzyme which method comprisesincubating the histone deacetylase enzyme with: (i) trypsin; (ii)acetyl-Gly-Ala-(N^(ε)-acetyllysine)-AMC; in the presence and absence ofthe test compound; and (iii) measuring the increase in fluorescence at460 nm over time, using an excitation wavelength of 355 nm wherein thedifference in the rate of increase of the fluorescence in the presenceand absence of the test compound is indicative of inhibitory propertiesof the test compound.
 19. The method of claim 18 where the histonedeacetylase enzyme is HDAC1.
 20. The method of claim 19 wherein HDAC1 isincubated with the test compound for at least 5 minutes prior toaddition of trypsin and acetyl-Gly-Ala-(N^(ε)-acetyllysine)-AMC.